Hypha-specific factors from candida albicans

ABSTRACT

The present invention relates to biochips, in particular nucleotide chips, which contain hyphen-specific proteins coding nucleotides, protein chips which contain hyphen-specific proteins, and antibody chips which contain antibodies directed against these hyphen-specific proteins, diagnostic compositions which contain these nucleotide, protein, or antibody chips, processes for the location and identification of substances which are therapeutically effective against diseases caused by types of Candida and processes for the diagnosis of a disease caused by Candida.

[0001] The present application is a nationalization of PCT ApplicationNo. PCT/EP01/05363, filed May 10, 2001, claiming priority to GermanPatent Application 10023130.6, filed May 11, 2000.

BACKGROUND

[0002] The present invention relates to biochips, in particularnucleotide chips, which contain nucleotide sequences codinghyphen-specific proteins, protein chips, which contain hyphen-specificproteins, and antibody chips, which contain antibodies directed againstthese hyphen-specific proteins, diagnostic compositions which containthese nucleotide, protein, or antibody chips, processes for the locationand identification of substances, which are therapeutically effectiveagainst diseases caused by types of Candida and processes for thediagnosis of a disease caused by Candida.

[0003] Along with the yeasts of the Saccharomycetaceae family, whichhave been used commercially, e. g. in the food industry, for a longtime, asporogenous yeasts such as, for example, yeasts of the genusCandida, also number among the budding funguses or yeasts. Severalmembers of the genus Candida are able to form mycel aggregates. Othersreproduce only by sprouting. Candida albicans is the most frequentlyisolated human pathogenic fungus. Candida albicans frequently causesopportunistic infections, i. e. infections in immunosuppressed patientsby normally relatively unproblematic microbes. Infections of this typetake a serious course in these patients and decisively shorten thesurvival time, for example, of HIV-infected patients or of cancerpatients treated with chemotherapy or radiation therapy. Presently, thetreatment of systemic infections with Candida albicans is carried outprincipally by means of azoles or polyenes. However, the treatment bymeans of these two classes of substances has disadvantages. Polyeneslead to strong side effects while resistance to the azoles isincreasingly developing (DiDomenico, 1999, Curr Opin Microbiol 2, 509 to515, Georgopapadakou, 1998, Curr Opin Microbiol 1, 547 to 557).

[0004] Since the clinical findings of fungus infections arepredominantly uncharacteristic, forming the exact diagnosis of fungalinfections, in particular of Candida infections, is extremely difficult.Where a Candida attack on the skin or the mucus membranes is suspected,for example, surface smears must be taken and examined microscopically.In attacks on internal organs, organ biopsies must be examinedhistologically in order to detect invasive growth. For the diagnosis ofa generalized Candida infection, a specular examination of the ocularfundus is indicated. Furthermore, several blood cultures, which must betaken venously on successive days, must be examined. In case the kidneysare involved the urine must be examined in addition. Microscopic nativepreparations of this kind permit, however, only the detection ofpolymorphic fungal cells (hyphens, pseudohyphens, and blastospores) andspores, indeed without enabling the exact species to be determined andspecific therapeutic measures to be introduced.

[0005] Along with the microscopic detection, it is thereforeindispensable to establish cultures for the determination of the exactspecies. A further diagnostic possibility, which nevertheless up to nowhas not had the desired informational value, is the detection of Candidaantigens in the serum of the patient. Although a high titer indeedargues for a systemic Candida infection, it still does not prove it,while a negative finding cannot rule out a systemic infection.

[0006] The development of further improved diagnostics for doubt-freeassignment of a disturbance of health to the infections caused byrepresentatives of the Candida family and of antimycotics for thetreatment of infections caused by representatives of the Candida familyis therefore urgently needed.

SUMMARY OF THE INVENTION

[0007] The technical problem underlying the present invention thereforeconsists of providing for the means and processes for the diagnosis ofinfections caused by Candida albicans and for the development ofsubstances, which are therapeutically effective against diseases causedby Candida. The invention solves the technical problem underlying itthrough the preparation of biochips, in particular, of a nucleotidechip, including a solid substrate and at least one nucleotide sequencefixed on it which is suitable for the identification and transcriptionof a gene coding for a hyphen-specific protein from Candida, inparticular Candida albicans, where this nucleotide sequence is selectedfrom the group consisting of:

[0008] (a) a nucleotide sequence, defined in SEQ ID no. 1, 2, 3, 4, 12,13, 15 or 17, or a complementary strand or portion thereof,

[0009] (b) a nucleotide sequence, coding an amino acid sequence, definedin SEQ ID no. 5, 6, 7, 8, 14, 16 or 18, or a complementary strand orpart thereof, and

[0010] (c) a nucleotide sequence, which hybridizes with one of thenucleotide sequences named in (a) or (b).

BRIEF DESCRIPTION OF THE DRAWINGS:

[0011] The invention will now be described, in association with thedrawings, in which:

[0012]FIG. 1 shows microscopic pictures of the virulent, hyphallygrowing Candida albicans strain Sc5315 and the non-virulent Candidastrain Can34 (Δcph1Δefg1) growing in the manner of a yeast;

[0013]FIG. 2a shows microscopic pictures of the virulent, hyphallygrowing Candida albicans strain Sc5315 and the non-virulent Candidastrain Can34 (Δcph1Δefg1) growing in the manner of a yeast;

[0014]FIG. 2b show the results of a Northern blot analysis with the useof RNAs from these two strains which previously were cultivated eitherYPD medium or in α-MEM medium; and

[0015]FIG. 3 shows the results of 2D gel electrophoreses of proteinextract from the strains Sc5315 and Can34 (Δcph1Δefg1) cultivated inα-MEM medium.

[0016] In connection with the present invention, it is understood that abiochip means a device which contains, in immobilized or fixed form, aplurality of biological substances—by way of example, nucleotidesequences, proteins or antibodies—and, with whose aid, a small quantityof a ligand, which under suitable circumstances can bind to thebiological substance, can be detected in a small sample by means ofhybridization and/or binding methods. It is understood that a nucleotidechip means a device which contains a plurality of different nucleicacids or nucleotide sequences such as DNA or RNA in immobilized form,and with whose aid a small amount of a complementary nucleic acid in asmall sample liquid or—by means of DNA/protein binding tests—a smallamount of a protein binding to nucleic acids can be detected.

[0017] According to the invention, the nucleotide chips contain, fixedon a solid substrate, nucleotide sequences, which exclusively codeproteins, which are expressed in the hyphally growing form of Candidaalbicans or which exclusively regulate the expression of hyphen-specificproteins. That is, the nucleotide sequences contained on the nucleotidechips according to the invention are not expressed during the yeast-likegrowth of Candida. The nucleotide sequences described according to theinvention and the proteins coded by them have no significant homologiesto a related, non-pathogenic, non-hyphally growing fungus, for examplewith proteins of Saccharomyces cerevisiae. It is known that thefilamentous growth, that is the formation of hyphae, is an importantprerequisite for the occurrence of the virulent qualities of Candida(Mitchell, 1998, Curr. Opin. Microbiol., 1, 687-692). Thus, forms ofCandida albicans, which form no hyphae, are non-virulent in a modelsystem (Mus musculus) (Lo et al., 1997, Cell 90, 939 to 949). Thus, inconnection with the present invention, it is understood that ahyphen-specific protein means a protein and/or peptide which isexpressed exclusively in the types of the genus Candida and preferablyhas significance for the virulence of Candida, in particular Candidaalbicans.

[0018] The nucleotide sequences used according to the invention,occurring specifically in the pathogenic form of Candida albicans andthe proteins coded by these, thus represent outstanding diagnosticauxiliary means for the recognition of local or systematic candidoses,in particular for the recognition of local or systemic Candida albicansinfections. Moreover, they offer the possibility, in the case of aCandida infection, of distinguishing hyphally growing, thereforevirulent, Candida albicans forms from yeast-like growing, thereforenon-virulent, forms of Candida albicans.

[0019] Moreover, the nucleotide sequences and proteins used according tothe invention prove to be particularly valuable for the development ofmedicaments for fighting candidoses. The nucleotide sequences andproteins according to the invention can be used as targets for theidentification specifically of substances effective against these. Thus,substance libraries can be searched for the interaction of substancespresent in them with proteins according to the invention or nucleotidesequences according to the invention.

[0020] The invention thus relates in its preferred form of embodiment toan aforementioned nucleotide chip, which includes a nucleotide sequence,which is a protein-coding nucleotide sequence, selected from the groupconsisting of the nucleotide sequences of SEQ ID No. 1, 2, 3, 4, 13, 15,and 17. These nucleotide sequences code the amino acid sequencesdescribed in SEQ ID No. 5, 6, 7, 8, 14, 16, and 18. These sequences areparticularly helpful in the diagnosis of diseases, which are caused bytypes of Candida, in that they enable the selective detection of thepresence of Candida in surface smears or organ biopsies. For example,the nucleotide sequences according to the invention contained on such anucleotide chip can be hybridized with marked DNA samples, which areisolated from sources such as skin smears, biopsy samples, or speciallyestablished fungal cultures, or amplified by means of PCR methods understringent conditions. Since the nucleotide sequences according to theinvention have no significant homologies to nucleotide sequences fromrelated funguses, the detection of a hybridization thus permits thedetection of Candida in a sample infected with fungus. The nucleotidesequences used according to the invention, however, make possible notonly the simple detection of Candida, but also the detection thatCandida is growing in hyphalic form and has the correspondingly virulentproperties. For example, mRNA can be selectively isolated from skinswabs or biopsy samples and/or amplified by means of PCR methods. Aftermarking with suitable marking materials, the mRNA so obtained ishybridized with the nucleotide chip containing the nucleotide sequencesaccording to the invention. Since the nucleotide sequences according tothe invention are expressed and transcribed exclusively during thehyphalic growth of Candida, not during the yeast-like growth, thedetection of a hybridization permits, with the use of isolated mRNA, thedetection that Candida is found in the transition to the hyphalic, andthus virulent, growth phase.

[0021] In an additional preferred form of embodiment, the inventionrelates to the aforementioned nucleotide chip, which contains nucleotidesequences which are regulator elements of hyphen-specific Candidaprotein-coding genes, therefore, elements, which in particular makepossible the transcription of the protein-coding regions functionallyconnected with these regulatory elements, for example promoters,transcription termination signals, silencers, enhancers, and so on.These particularly preferred nucleotide sequences can in particular bepromoters, particularly preferably the promoter represented in SEQ IDNo. 12. The regulatory elements according to the invention, inparticular promoters, particularly preferably the promoter described inSEQ ID. No. 12, prove to be particularly advantageous to the extent thatthey include the regulation sequences necessary for the induction ofhyphen-specifically expressed proteins and accordingly can be used inorder to identify additional specific Candida proteins which, throughbinding to these regulatory elements, can induce or inhibit in vivo thetranscription of hyphen-specific proteins, in particular of proteinswith the amino acid sequences described in SEQ ID No. 5, 6, 7, 8, 14,16, and 18. After the identification of such proteins, nucleotide chips,which contain the regulatory elements of hyphen-specifically expressedprotein-coding nucleotide sequences can be used to identify substancesof any kind, which inhibit the interaction between the regulatoryelements and the proteins binding thereto. With the use of nucleotidechips of this type it is therefore possible to identify substances,which inhibit the expression of hyphen-specific proteins and that thuscan potentially be used as medicaments acting specifically againstCandida infections.

[0022] The invention relates, in a further particularly preferred formof embodiment, to the aforementioned nucleotide chip, which has DNA,RNA, or PNA sequences as nucleotide sequence. PNA (peptide nucleic acid,or polyamide nucleic acid) sequences are molecules, which are notnegatively charged and act in a manner similar to DNA (Nielsen et al.,1991, Science, 254, 1497-1500; Nielsen et al., 1997, Biochemistry, 36,5072-5077; Weiler et al., 1997, Nuc. Acids Res., 25, 2792-2799). PNAsequences include a polyamide backbone of N-(2-aminoethyl)-glycineunits, and have no glucose units and no phosphate groups.

[0023] According to the invention, the nucleic acid molecules, which canbe fixed on a substrate, can be isolated from natural sources,preferably from Candida albicans. For example, the nucleic acidmolecules can be isolated and amplified by means of PCR methods, wherebydouble-stranded molecules are obtained. According to the invention, thenucleic acid molecules can, however, also be synthesized in vitroaccording to known processes, whereby single-stranded oligonucleotidesor peptide-oligonucleotides are obtained. Through the choice of asuitable primer, desired regions of the nucleic acids, according to theinvention, i. e. individual regions as well as the whole reading frameof the gene, can be isolated and amplified. By means of currentmolecular biological techniques it is possible to introduce mutations ofvarious kinds into the nucleic acid molecules, according to theinvention. Thereby, for example, sequence variants can be realized,which appear in different clinical Candida isolates. These types ofmutations realized by the invention can be insertions, deletions,duplications, inversions, additions, exchanges or the like, even ofunusual nucleotides. In this way, however, it is also possible toproduce modified oligonucleotides with functional groups, which makepossible a covalent binding of the oligonucleotide to the substratematerial for the production of the nucleotide chips according to theinvention. Thus, for example, oligonucleotides with amino modificationsor biotin groups can be produced, which can bind covalently tochemically reactive groups (epoxides) contained on the surface of thesubstrate material or to streptavidin groups or derivatives thereof. Ina further preferred form of embodiment of the invention, it is providedthat nucleic acids are provided with nucleoside derivatives containingphotosensitive protective groups.

[0024] According to the invention, nucleotide sequences can also be usedfor the nucleotide chip, which are produced by fusion of the nucleotidesequences according to the invention with genes or segments of genesfrom other sources. According to the invention, shortened nucleotidesequences of the aforementioned type can also be used, to the extentthat they have said hyphen specificity. According to the invention, itis provided that shortened nucleotide sequences have a length of atleast 15 base pairs.

[0025] In an additional form of embodiment of the invention, it isprovided that the nucleotide chip also contains nucleotide sequences ofthe aforementioned type, which according to the invention, hybridizewith the aforementioned nucleotide sequences. Hybridization inconnection with this aspect of the invention means hybridization underconventional hybridization requirements, as they are described inSambrook et al. (Molecular Cloning. A Laboratory Manual. Cold SpringHarbor Laboratory Press, 2nd edition 1989), preferably under stringentrequirements. According to the present invention, one speaks of ahybridization if after washing for an hour with 1 ×SSC and 0.1% SDS at55° C., preferably at 62° C., and particularly preferably at 68° C., inparticular for one hour [with] 0.2 ×SSC and 0.1% SDS at 55° C.,preferably at 62° C., and particularly preferably at 68° C., a positivehybridization signal is still observed. According to the invention, anucleotide sequence hybridizing with one of the nucleotide sequencesspecified in the sequence protocols can, under washing conditions ofthis type, be used, by immobilization on the solid substrate, for thenucleotide chip, according to the invention.

[0026] The identification and isolation of hybridizing nucleotidesequences can be done, by way of example, with the use of one of thenucleotide chips, according to the invention, said nucleotide chipcontaining the aforementioned nucleotide sequences or parts of thesemolecules or their complementary strands. The nucleotide chip used forthe identification and isolation of hybridizing nucleotide sequences cancontain, by way of example, nucleotide sequences which have exactly oressentially the nucleotide sequences represented under SEQ ID No. 1 to4, 12, 13, 15 or 17, or parts of these sequences or complementarystrands. However, the nucleotide chip can also contain syntheticfragments, which have been produced with the aid of the usual synthesistechniques and whose sequence, according to the invention, agreesessentially with one of the nucleotide sequences. In this way,nucleotide sequences from clinical Candida isolates, according to theinvention, can be isolated and be made available for the nucleotidechip, which contain deviations or mutations with respect to thenucleotide sequences described in SEQ ID No. 1 to 4,9to 13, 15 or 17.

[0027] According to the invention, the molecules hybridizing with thenucleotide sequences also include fragments, derivatives, functionalequivalents and/or allelic variants of the above-described nucleotidesequences, which code a protein according to the invention or guaranteeits hyphen-specific expression. In the statement above “fragments” areunderstood to mean parts of the nucleotide sequences, which are longenough in order to code the hyphen-specifically expressed protein or toassure hyphen specificity. The expression “derivative,” “functionalequivalent,” or “mutant deviation” means in connection with the presentinvention that the sequences of these molecules differ from thesequences of the above-described nucleotide sequences in one or morepositions, but have a high degree of homology to these sequences at thenucleotide level. Homology here means a sequence identity of at least40%, in particular an identity of at least 60%, preferably over 80% andparticularly preferably over 90%, 95%, 97% or 99% at the nucleic acidlevel.

[0028] According to the invention, the nucleotide chips containnucleotide sequences fixed or immobilized on a solid substrate. Inconnection with the present invention, the term “solid substrate” meansan insoluble matrix. In the preferred form of embodiment, the solidsubstrate consists of a hydrophobic or weakly hydrophilic material liketransparent glass, silica dioxide, metal oxides, polymers and copolymersof dextrans or amides, by way of example, acryl amide derivatives,cellulose, nylon, or polymeric materials such as polyethyleneterephthalate, cellulose acetate, polystyrene or polymethylmethacrylate,or a polycarbonate of bisphenol A. Before fixing the nucleotidesequences, the substrate material is preferably pretreated with asurface-activating agent such as poly-L-lysine, polyethylenimine, orpolyalkylamine in order to improve the fixing of the nucleotidesequences on the substrate material. In another form of embodiment,glass used as substrate is pretreated with a silane-coupling agent,which has an amino group, an aldehyde group, or an epoxy group. However,the commercially available precoated types of substrate such aspoly-L-lysine (Sigma Diagnostics), Super-Aldehyde (Telechem),Super-Amine (Telechem), Silane Prep (Sigma), CMT GAPS (Corning), Type I(Clontech), Type II (Clontech), Arraylink (GeneScan Europe), Type I(Eppendorf), Type II (Eppendorf), Epoxysilan (Quantifoil) andCast/FastSlides (Schleicher & Schüll) can also be used for thenucleotide chip according to the invention. Other suitable substratesare those that are used for photolithographically produced nucleotidechips, for example, those described in Lipshutz et al. (Lipshutz, Fodor,Gingeras and Lockhart, 1999, Nat. Genet., 21, 20-24). In a particularlypreferred form of embodiment, substrates are used which have a coatingof poly-L-lysine, as described in DeRisi et al. (DeRisi, J. L., Iyer, V.R., and Brown, P. O., 1997, Science, 278, 680-686), for examplePoly-Prep slides (Sigma Diagnostics), or aminosilanes, like Silane-Prepslides (Sigma), CMT GAPS slides (Corning), and Super Amine (Telechem),or membranes, like CAST slides or FAST slides (Schleicher & Schüll). Forimmobilization of amino-modified oligonucleotides, epoxy-modifiedsurfaces, like ArrayLink Biochip (GeneScan Europe) or Epoxysilane slides(Quantifoil) are particularly preferred.

[0029] The nucleotide sequences can be bound and fixed on the carriersubstrate via chemical or photochemical reactions or by electrostaticinteractions. In a preferred form of embodiment of the invention, it isprovided that the immobilization or fixing of the nucleic acid on thesubstrate surfaces used is accomplished via an electrostatic or acovalent binding. If the nucleic acids, for example, were producedsynthetically and have a functional group, the nucleic acids can bebound covalently and fixed on the surface of the substrate material.(Lamture et al., 1994, Nucl. Acids Res., 22, 2121-2125; Guo et al.,1994, Nucl. Acids Res., 22, 5456-5465). According to the invention, thenucleic acids can also be covalently bound to the surface-activatedsubstrate via a spacer or a cross-linking agent, for example, abifunctional cross-linking agent. In a preferred form or embodiment ofthe invention it is provided that the binding of the nucleotidesequences on the substrate is done, in the case of polylysine-coated,aminosilane-coated, and membrane-coated nucleotide chips, by means of UVcross linking and, in case of epoxy modified chips, by means of achemical reaction. The binding of the nucleic acids on the substrate canobviously also be done via photochemical reactions. In the case of suchphotolithographically produced nucleotide chips, a selective splittingoff of the photosensitive protective group occurs by means ofphotolysis.

[0030] A further preferred form of execution of the present inventionthus relates to processes for the production of nucleotide chipsaccording to claim 1, including the isolation and/or amplification of atleast one nucleotide sequence, which codes a hyphen-specificallyexpressed protein of Candida and/or contains regulatory elements of thisnucleotide sequence, or the chemical synthesis of this nucleotidesequence, the modification of the nucleotide sequence during or afterthe synthesis or amplification by the insertion of functional groups orspacer units, the application of an aqueous solution of the isolated orsynthesized nucleotide sequence to a solid substrate material, and theimmobilization of the nucleotide sequence on the substrate by means ofchemical or photochemical reaction or electrostatic interaction.

[0031] A further particularly preferred form of execution of theinvention, relates to protein chips containing a solid substrate and atleast one hyphen-specific protein fixed on it, selected from the groupconsisting of:

[0032] (a) a protein with an amino acid sequence defined in SEQ ID No.5, 6, 7, 8, 14, 16 or 18, or a fragment thereof, and

[0033] (b) a protein with an amino acid sequence which has a sequenceidentity of at least 80% to one of the amino acid sequences defined inSEQ ID No. 5, 6, 7, 8, 14, 16 or 18, or to a fragment thereof.

[0034] In connection with the present invention, a protein chip isunderstood to mean a device which contains a plurality of differentproteins or peptides in immobilized form and, with whose aid, a smallamount of a ligand, e. g. of a protein or an antibody which can bindcovalently or non-covalently to at least one protein or peptide fixed tothe substrate, can be detected in a small sample liquid.

[0035] According to the invention, the protein chips contain proteinsfixed on a solid substrate, said proteins being expressed only in thehyphally growing form of Candida albicans, or parts thereof. The proteinchips according to the invention can thus, for example, be used for thedetection of antibodies, which were formed in the body of an organism,in particular a mammal as a consequence of an immunization by antigendeterminants of hyphally growing forms of Candida, in particularhyphen-specific Candida proteins. The binding of at least one antibodyfor blood, lymph, body secretions, or other body fluids of an organismto the protein chip according to the invention therefore makes possiblethe detection of a systemic Candida infection in this organism, saidinfection leading to the formation of the bound antibody. Moreover, theprotein chips according to the invention can also be used in order, forexample, to identify and isolate such proteins from Candida-infectedmaterials as enter, in vivo, into interactions with the proteinscontained on the protein chip. After identification and isolation ofinteracting proteins of this type, the protein chips, according to theinvention, can also be used in order to identify substances of any type,which according to the invention, can inhibit or promote the interactionbetween the proteins and proteins interacting therewith. With the use ofthe protein chips according to the invention therefore, substances canbe detected which are potentially suitable as medicaments for fightingCandida infections, in particular for the inhibition of the transitionfrom yeast-like growth to hyphal growth of Candida.

[0036] In a particularly preferred form of embodiment of the invention,it is provided that the protein chip, according to the invention, canalso include, along with the hyphen-specific proteins with the aminoacid sequences represented in SEQ ID 5, 6, 7, 8, 14, 16, and 18, alsoderivatives, functional equivalents, or variants of these proteins. Inconnection with the present invention the phrase “derivatives,functional equivalents, or variants” is understood to mean, inparticular, those derivatives of the proteins with the amino acidsequences specified in SEQ ID 5 to 8, 14, 16, and 18 which, whileretaining the basic structure of these proteins, are obtained bysubstitution of atoms or molecule groups and their amino acid sequencesare distinguished from the amino acid sequences specified in SEQ ID 5 to8, 14, 16, and 18 at least one position and which essentially have ahigh degree of homology at the amino acid level. The term “homology”known to those skilled in the art denotes the degree of kinship betweentwo polypeptides, said degree being determined by the extent ofagreement between these polypeptides. Therein an agreement can mean anidentical agreement, thus sequence identity, as well as a conservativeamino acid exchange. Preferably derivatives, variants, or functionalequivalents used according to the invention each have a sequenceidentity to one [of the] amino acid sequences specified in SEQ ID 5 to8, 14, 16, and 18 of at least 80%, preferably 85%, and particularlypreferably of over 90%, 95%, 97%, and 99% at the amino acid level. Thedeviations relative to the amino acid sequences represented in SEQ ID 5,6, 7, 8, 14, 16 or 18 could have arisen, for example, by deletions,substitutions, insertions, additions, exchanges, or recombinations ofthe nucleotide sequences coding the amino acid sequences produced withtechnological means. However, they can be naturally occurringvariations, for example, amino acid sequence changes arising in anatural manner. Derivatives or variants of the hyphen-specific proteinsaccording to the invention can, for example, stem from clinical isolatesof Candida.

[0037] Such derivatives, functional equivalents, or variants can bedistinguished from the proteins with the amino acid sequencesrepresented in SEQ ID 5, 6, 7, 8, 14, 16, and 18, for example, by analtered stability, specificity, a modified temperature, pH value, and/orconcentration profile, an altered activity, and/or an altered effectorpattern. Derivatives, functional equivalents, or variants can also occurin other conformations or have other subunits or pre-translationaland/or post-translational modifications. Despite the differences whichmay be present, the hyphen-specific proteins with the amino acidsequences represented in SEQ ID 5, 6, 7, 8, 14, 16, and 18 andderivatives, variants, or functional equivalents thereof havenonetheless certain common characteristics such as activity, molecularweight, immunological reactivity, conformation, and/or physicalproperties such as running behavior in gel electrophoresis and theirsolubility and other things.

[0038] In an additional particularly preferred form of embodiment of theinvention, it is provided that the protein chip according to theinvention includes fragments of the proteins with the amino acidsequences specified in SEQ ID 5 to 8, 14, 16 or 18 or the proteins whoseamino acid sequence has a sequence identity of at least 80% relative toone of the amino acid sequences defined in SEQ ID 5, 6, 7, 8, 14, 16,and 18. In connection with the present invention, “fragments” isunderstood to mean, in particular those isolated regions of a protein,which have fewer amino acids than the native protein but whose length issufficient that the isolated fragment can exert one of the functionscharacteristic for the native protein such as binding capability to asecond protein, a specific catalytic activity, and so on. In aparticularly preferred form of embodiment of the invention, the fragmentof a protein includes a protein region, which represents an antigendeterminant or an epitope and thus is suitable to a particular degreefor the binding of an antibody.

[0039] According to the invention, the hyphen-specific proteins, inparticular the proteins with the amino acid sequences represented in SEQID 5, 6, 7, 8, 14, 16, and 18, which according to the invention, areimmobilized on the protein chip, can have been isolated and purifiedfrom natural sources, for example, from Candida-infected tissues orspecifically established Candida cultures with the use of customaryprocesses known in the art. The proteins or fragments used can also beof synthetic origin. For example, peptides, that is fragments of theproteins according to the invention, can be produced with the aid of theprocess of Merrifield (1985, Angew. Chem. 97, 801). In a particularlypreferred form of embodiment of the present invention thehyphen-specific proteins or peptides are produced by means of customaryDNA recombination technologies. For example, the protein-codingnucleotide sequences according to the invention such as the sequencesrepresented in SEQ ID 1, 2, 3, 4, 13, 15, and 17, or sequenceshybridized therewith are inserted in suitable vectors with the use ofcustomary processes of molecular biology and gene technology and cloned.Preferably, the insertion of the protein-coding nucleotide sequences,according to the invention, is done so that they are under the controlof regulatory elements, i. e. are operationally connected with them.These regulatory elements assure the transcription and synthesis oftranslatable nucleic acid molecules in prokaryotic and eukaryotic cells.The regulatory elements can be promoters, enhancers, operators,silencers, and/or transcription termination signals. Aftertransformation in suitable host cells, for example prokaryotic andeukaryotic cells such as bacteria, yeast, plant, insect, or mammalcells, these host cells can be cultivated in a suitable culture mediumunder such conditions as permit the formation of the hyphen-specificproteins coded by the protein-coding nucleotide sequence or a fragmentthereof. Subsequently, the protein or fragment thereof can, with the useof a suitable process, be isolated from the host cell, or the medium inwhich the host cell was cultivated, and purified. However, for theproduction of the hyphen-specific proteins or fragments suitable invitro transcription/translation systems can also be used. In aparticularly preferred form of embodiment of the invention, theproduction of the hyphen-specific proteins or fragments thereof is donein bacterial expression systems wherein the protein preferably areobtained as GST fusion proteins, HIS-tag fusion proteins, pMAL fusionproteins, and so on.

[0040] As a solid substrate for the protein chips according to theinvention, the same materials can be used as described above for thenucleotide chips according to the invention, for example glass, silicondioxide, other silica materials, polymeric materials such asfluoropolymers, metal oxides, etc. can be used. These substratematerials are preferably pretreated before immobilization of theantibodies, for example, with silane coupling agents. In a preferredform of embodiment of the invention, the epoxy-modified substrate ormembranes described by Joos et al. (Joos et al., 2000, Electrophoresis,21, 2641-2650) are used.

[0041] According to the invention, it is provided that the binding andimmobilization of the hyphen-specific proteins or fragments on thesubstrate material is done by means of chemical or photochemicalreaction or electrostatic interaction. The hyphen-specific proteins,according to the invention and fragments thereof can, for example, bebound and immobilized on the substrate material by a plurality ofcustomarily used functional groups and/or spacers or chemicalcross-linking agents, such as bifunctional cross-linking agents. Anoverview of suitable functional groups which make possible a binding ofproteins to silanized surfaces is found, for example, in Weetall, 1996,Advances in Molecular and Cell Biology, Vol. 15A, 161-192, JAI PressInc. In case the protein to be immobilized is present as GST fusionprotein, the binding of the protein to the substrate is done via GSHunits present on the surface of the substrate. The immobilization of apMAL fusion protein can be done via MBP units on the surface of thesubstrate. If the protein to be immobilized is HIS-tag fusion protein,the immobilization can be done via Ni²⁺-nitrilotriacetic acid surfaces(Ni-NTA) (Adachi, et al., Proc. Nat. Acad. Sci. USA, 97, 7243-7247).

[0042] An additional preferred form of embodiment of the presentinvention thus relates to processes for the production of protein chipsincluding the isolation of at least one hyphen-specifically expressedCandida protein from a suitable source or the chemical synthesis orrecombinant production of this protein or a fragment thereof, themodification of the protein or fragment during or after the isolation,synthesis or production by the insertion of functional groups or spacerunits, the application of an aqueous solution of the isolated orsynthesized protein on a solid substrate material by means of chemicalor photochemical reaction or electrostatic interaction.

[0043] A particularly preferred form of embodiment of the presentinvention, relates to an antibody chip including a solid substrate andat least one antibody fixed thereon which is directed specificallyagainst a protein with the amino acid sequence represented in SEQ ID 5,6, 7, 8, 14, 16 or 18, or a fragment thereof or a derivative thereof.

[0044] Since the antibodies fixed on the antibody chip, according to theinvention, are directed specifically against hyphen-specific Candidaproteins, the presence of hyphally growing Candida cells in sections ofskin or mucus membrane infected by fungus, organ biopsies, or bodyfluids can be detected with the use of a chip of this type. For example,proteins can be extracted from the aforementioned samples and, aftermarking with the antibody chip, according to the invention, incubated.The binding of at least one marked protein on the antibody chip,according to the invention, thus shows that hyphally growing Candida ispresent in the sample examined.

[0045] In connection with the present invention, the term “antibody” isunderstood to mean a polypeptide, which is coded by one or moreimmunoglobin genes and recognizes specific structures on an antigen, inparticular an antigen determinant or an epitope, and can bindspecifically thereto. The term “antibody” does not include only acomplete immunoglobin but rather also a series of fragments, which areavailable by means of splitting with various peptidases. The term“antibody” also includes modified antibodies such as oligomers, reduced,oxidized, and marked antibodies. “Antibody” also includes antibodyfragments, which were produced by modification of intact antibodies aswell as with the use of DNA recombination technologies. In connectionwith the present invention “antibody” also includes, in particular,fragments such as Fab, F(ab′)₂, or Fvm, which can bind to an antigendeterminant. The Fab fragment can be produced by splitting of the intactantibody with the enzyme papain whereby an intact light chain with apart of a heavy chain is obtained. F(ab′)₂ can be produced by treatmentof the intact antibody with pepsin without subsequent reduction. F(ab′)₂is a dimer consisting of two Fab′ fragments. Fv is an antibody fragmentproduced by gene technology, said antibody fragment including thevariable region of the light chain and the variable region of the heavychain. Processes for the production of such fragments have beendescribed, for example, by Harlow and Lane in “Antibodies: A LaboratoryManual”, 1988, Cold Spring Harbor Laboratory, New York.

[0046] The phrase “antibody which is directed against a protein” or“antibody, which binds specifically to a protein” means that an antibodycan recognize, under defined immune test conditions, an antigendeterminant or an epitope of a protein and can bind thereto by means ofits paratope. Antigen determinants usually consist of chemically activemolecule groups such as amino acids or sugar side chains on the surfaceof an antigen, e. g. a protein, and have a characteristicthree-dimensional structure. Under defined conditions an antibody thuspreferably binds only to a certain protein while no noteworthy bindingto other proteins in the same sample occurs.

[0047] An antibody immobilized according to the invention on an antibodychip can thus bind to a protein, peptide, carbohydrate, proteoglycan,and/or a lipid complex, which is in specific relation with thehyphen-specific protein used according to the invention. According tothe invention, an antibody used can be directed against structures,which are to be considered as post-translational modifications of thehyphen-specific proteins.

[0048] According to the invention, it is provided that the antibody chipcontains monoclonal as well as polyclonal antibodies, which are able tospecifically identify, and in given cases bind to, a structure of ahyphen-specific protein, according to the invention.

[0049] According to the invention, the monoclonal and polyclonalantibodies contained on the antibody chip can be produced and isolatedwith the use of processes well-known in the art. The processes for theproduction of monoclonal antibodies with the use of hybridoma technologyare described, for example, in “Hybridoma Techniques” (1980) or in theU.S. Pat. No. 4,341,761, U.S. Pat. No. 4,399,121, or U.S. Pat. No.4,472,500.

[0050] As substrate material for the immobilization of the antibodies,the materials named above for protein chips such as glass, silicondioxide, nylon, acryl amide derivatives, silica materials, polymericmaterials, such as fluoropolymers, metal oxides, etc. can be used. Thesesubstrate materials are preferably pretreated before immobilization ofthe antibodies, for example, with silane coupling agents. In a preferredform of embodiment of the invention, the epoxy-modified substrate ormembranes described by Joos et al. (Joos et al., 2000, Electrophoresis,21, 2641-2650) are used.

[0051] According to the invention, it is provided that the binding ofthe antibodies, directed against specific hyphen-specific proteins, onthe substrate is done via a chemical or photochemical reaction or viaelectrostatic interactions. As described above for protein chips, theantibodies used, according to the invention, e. g. with the aid of aplurality of customarily used functional groups and/or spacers orchemical cross-linking agents such as such as bifunctional cross-linkingagents, are bound and immobilized on the substrate material.

[0052] An additional preferred form of embodiment of the presentinvention, thus relates to processes for the production of antibody hipsincluding the gene-technological production, isolation, or synthesis ofat least one antibody directed against a hyphen-specific Candida proteinor fragments thereof, the modification of the antibody or fragmentthereof during or after the isolation, synthesis, or production by theinsertion of functional groups or spacer units, the application of anaqueous solution of the isolated or synthesized protein to a solidsubstrate material by means of chemical or photochemical reaction orelectrostatic interaction.

[0053] An additional particularly preferred form of embodiment of thepresent invention, relates to an antibody chip including a solidsubstrate and at least one antibody fixed thereon which is directedspecifically against a protein with the amino acid sequence representedin SEQ ID 5, 6, 7, 8, 14, 16 or 18. An antibody chip of this typetherefore includes antibodies, which are directed against theaforementioned antibody and can recognize and bind to it specifically.The use of an antibody makes possible the detection of antibodiesagainst hyphen-specific proteins of Candida in the blood, the lymph, inbody secretions, or other body fluids of an organism and thus thedetection of a systemic Candida infection in this organism, which haslead to the formation of the antibody contained in the sample.

[0054] The present invention also relates to a diagnostic compositionincluding at least one nucleotide chip, according to the invention, aprotein chip, according to the invention and/or an antibody chip,according to the invention. The invention, thus, also includesdiagnostic kits which, according to the invention, contain the biochipsi. e. nucleotide chips, protein chips, and antibody chips, suitablebuffer systems, and suitable marking systems.

[0055] The invention relates in a particularly preferred form ofembodiment to processes for the diagnosis of diseases caused by types ofCandida, in particular diseases caused by Candida albicans wherein asample to be tested is brought, in a suitable medium, into contact witha nucleotide chip according to the invention, a protein chip accordingto the invention, and/or an antibody chip, according to the inventionand an interaction between the sample to be tested and at least one ofsaid biochips is detected. The process, according to the invention, forthe diagnosis of Candida diseases is based on the detection of thepresence of nucleotide sequences, proteins, antibodies, or fragmentsthereof which, according to the invention, are in association with thehyphen-specific proteins in samples such as skin or mucus membraneswabs, organ biopsies, body secretions, blood, lymph, or other bodyfluids etc. which show fungal infection or are suspected of beinginfected.

[0056] In connection with the present invention, the phrase “diseases,disease states, or infections caused by types of Candida” is understoodto mean those diseases, which are causes exclusively by types ofCandida, in particular, however, by Candida albicans. The term thus alsoincludes all the diseases, which are caused by types of Candida such asC. tropicalis, C. krusei, C. parapsilosis, and C. guilliermondii, orTorulopsis (Candida) galabrata. According to the invention, this term isalso understood to include diseases or disease states which primarilyhave other causes and in which the types of Candida are only a part ofthe disease profile or additional symptoms are added, for example,opportunistic infections. The phrase “diseases, disease states, orinfections caused by types of Candida” includes, in particular, thosediseases such as Candida mycoses or candidoses, which can be subdividedessentially into three principal forms. The first principal form ofcandidosis is characterized by a saprophytic infestation of the skin andmucus membranes, in particular in the external genitalia, in the mouth,region of the nasal cavity, and in the digestive tract. The secondprincipal form of candidosis includes infections of the skin and mucusmembranes and is to a significant extent promoted by factors such aspregnancy, diabetes mellitus, serious diseases or traumas, cytostaticsand antibiotic therapy as well as alcoholism. The third principal formof candidosis includes deep organ mycoses in immune-suppressed patientswith cellular immune deficiency, in particular in the region of therespiratory tract, more rarely as Candida endocarditis, Candidameningitis, Candida nephritis, Candida endophthalmitis.

[0057] Based on the hyphen specificity and, associated therewith, thecorrelation to a Candida-caused disease profile, the presence ofnucleotide sequences, proteins, antibodies, or fragments thereof whichare in association with the hyphen-specific proteins according to theinvention points toward a Candida-caused disease. The detection of theaforementioned substances is done by binding and/or hybridization of atleast one of the biochips, according to the invention, whichspecifically recognize the substances to the detected.

[0058] If nucleotide sequences in association with the hyphen-specificproteins are detected in a sample, their detection, according to theinvention, is done by hybridization with the nucleotide chip. For this,nucleic acids, by way of example DNA or mRNA, are isolated from a sampleor from a specifically established culture and/or and amplified, forexample, by means of PCR processes. The nucleic acids extracted aresubsequently marked, for example, with fluorescent dyes, enzymes, orradioactive groups. If the extracted or marked DNA hybridizes with thenucleotide chip, then this shows the presence of Candida in the sampleexamined. If the extracted mRNA hybridizes with the nucleotide chipaccording to the invention, then this points to the fact that the samplecontains hyphally growing forms of Candida.

[0059] If hyphen-specific Candida proteins are to be detected in anexamined sample, proteins from the sample are extracted and marked andsubsequently incubated with the antibody chip according to theinvention, said antibody chip containing antibodies againsthyphen-specific proteins. According to the invention, the binding of atleast one marked protein on the antibody chip points to the presence ofhyphally growing Candida cells. According to the invention, thedetection of antibodies against Candida proteins in body fluids can bedone with the use of the protein chip as well as the antibody chip,which, according to the invention, contains the antibodies, which aredirected against the antibodies against hyphen-specific proteins.

[0060] The present invention also relates to processes for finding andidentifying substances, which are therapeutically effective againstCandida-caused diseases wherein a substance to be tested is brought, ina suitable medium, into contact with a nucleotide chip according to theinvention, a protein chip according to the invention, or an antibodychip according to the invention and an interaction between the sample tobe tested and at least one of said biochips is detected. Thus, accordingto the invention, the nucleotide chips and protein chips as describedabove, are used in order to identify substances, e. g. proteins, whichin vivo bind to nucleotide sequences, which code the hyphen-specificallyexpressed proteins or regulate the expression of these proteins or bindto the hyphen-specific proteins themselves. Such binding substances, inparticular proteins, can potentially be suitable as medicaments againstCandida-caused diseases if they, for example, are able, by binding toregulatory nucleotide sequences, to inhibit or restrict thetranscription of hyphen-specific proteins or if they are able, bybinding the hyphen-specific proteins, to inhibit or restrict theiractivity. If such substances, which, according to the invention, bind tothe nucleotide chips and protein chips induce or promote transcriptionof the hyphen-specific proteins or favor the activity of hyphen-specificproteins of this type of protein, the biochips, according to theinvention, can be used in order to identify additional substances whichare able to influence or inhibit the interaction between ahyphen-specific protein or the nucleotide sequence coding it and thesubstance binding thereto, in particular a binding protein. Suchsubstances are also potentially suitable as medicaments for thetreatment of Candida-caused diseases.

[0061] The present invention will be explained by the following sequenceprotocol and the following figures and examples.

[0062] The sequence protocol is part of this description and containsthe sequence protocols SEQ ID No. 1 to 18. Each of the amino acidsequences cited below was derived from the corresponding DNA sequenceand then partially verified by sequencing of the isolated proteins.

[0063] SEQ ID No. 1 represents the DNA sequence from Contig4-2149 codingCap33a.

[0064] SEQ ID No. 2 represents the DNA sequence from Contig4-2501 codingCap33b.

[0065] SEQ ID No. 3 represents the DNA sequence from Contig4-2069 codingCap18p.

[0066] SEQ ID No. 4 represents the DNA sequence from Contig4-2069 codingCap19p.

[0067] SEQ ID No. 5 represents the amino acid sequence of Cap33a.

[0068] SEQ ID No. 6 represents the amino acid sequence of Cap33b.

[0069] SEQ ID No. 7 represents the amino acid sequence of Cap18p.

[0070] SEQ ID No. 8 represents the amino acid sequence of Cap19p.

[0071] SEQ ID No. 9 represents the entire DNA sequence of Contig4-2149.

[0072] SEQ ID No. 10 represents the entire DNA sequence of Contig4-2501.

[0073] SEQ ID No. 11 represents the entire DNA sequence of Contig4-2069.

[0074] SEQ ID No. 12 represents the promoter region of Cap18p andCap19p.

[0075] SEQ ID No. 13 represents the DNA sequence from Contig5-3226coding Cap15p.

[0076] SEQ ID No. 14 represents the amino acid sequence of Cap15p.Cap15p is probably a nucleoside diphosphate kinase.

[0077] SEQ ID No. 15 represents the DNA sequence from Contig4-2178coding Cap20p.

[0078] SEQ ID No. 16 represents the amino acid sequence of Cap20p.Cap20p is probably a glutathione peroxidase.

[0079] SEQ ID No. 17 represents the DNA sequence from Contig5-2806coding Cap40p.

[0080] SEQ ID No. 18 represents the amino acid sequence of Cap40p.Cap40p is probably a fructose biphosphate aldolase.

[0081]FIG. 1 shows, in its left part, microscopic pictures of thevirulent, hyphally growing Candida albicans strain Sc5315 and thenon-virulent Candida strain Can34 (Δcph1Δefg1) growing in the manner ofa yeast. From both strains RNA was isolated and rewritten into cDNA. Themarked cDNA was hybridized with a nucleotide chip according to theinvention, on which nucleotide sequences coding CAP33, CAP19, and CAP18were fixed. The results of this hybridization are to be seen in theright part of FIG. 1. While the cDNA from the non-virulent strainhybridized with none of the nucleotide sequences contained on thenucleotide chip, the cDNA of the virulent strain showed stronghybridization signals with all three immobilized nucleotide sequences.If, however, the C. albicans strain Sc5315 is cultivated underconditions under which no hyphae arise, the same result is obtained asfor the non-virulent strain Can34 (Δcph1Δefg1).

[0082]FIG. 2a shows microscopic pictures of the virulent, hyphallygrowing Candida albicans strain Sc5315 and the non-virulent Candidastrain Can34 (Δcph1Δefg1) growing in the manner of a yeast. Under this,presentations of differential proteome analyses of the two strains aftercultivation in α-MEM medium are shown. From these presentations itfollows that Sc5314 on cultivation in α-MEM medium expresses theproteins p33a and p33b, but Can34 (Δcph1Δefg1) does not.

[0083]FIG. 2b show the results of a Northern blot analysis with the useof RNAs from these two strains which previously were cultivated eitherYPD medium or in α-MEM medium. Also included were RNAs from the virulentstrain Can16 (Δcph1) and the strain Can33 (Δefg1), which shows noformation of hyphae and has a sharply reduced virulence, where bothstrains were cultivated before RNA extraction in α-MEM medium. Onhybridization with cap18-specific, cap-19-specific [sic], andcap33-specific probes, hybridization signals were found with the RNAsisolated from the strains Sc5314 and Can16 cultivated in α-MEM medium.The RNA isolated from the strain Sc5314 cultivated in YPD medium showedon the contrary no hybridization signals. The RNA from the non-virulentstrain Can34 (cultivated either in YPD or in α-MEM medium) also showedno hybridization signals. The same result was also obtained in the RNA,which was isolated from the strain Can33 cultivated in α-MEM medium. Asa control an actin-specific probe (ACT1) was used.

[0084]FIG. 3 shows the results of 2D gel electrophoreses of proteinextract from the strains Sc5315 and Can34 (Δcph1Δefg1) cultivated inα-MEM medium. While Sc5315 expresses the hyphen-specific proteins p33a,p33b, p40, p15, p18, p19, and p20, these proteins are not expressed inCan34 (Δcph1Δefg1).

[0085] Additional advantageous developments of the invention aredescribed in the subordinate claims.

[0086] The invention will be described in more detail with the aid ofthe following examples.

EXAMPLE 1

[0087] The Isolation of the Proteins

[0088] The proteins were isolated from the clinical isolate Sc5314 bydifferential 2D gel electrophoresis as follows:

[0089] For the isolation of the proteins the virulent Candida albicansstrain Sc 5314 and the non-virulent Candida albicans strain Can34(HLC69) (Lo et al., 1997, Cell, 90, 939-949) were covered simultaneouslyin complete medium (YPD: 20 g/l bactopeptone, 10 g/l yeast extract, 0.15g/l L-tryptophan) over night, inoculated, in α-MEM medium (#22571 Lifetechnologies/Gibco), with 2% glucose extract (1:100), and incubated for24 hours at 37° C. on a rotary agitator. The cells thus obtained werepelletized and closed up in a saline buffer not containing detergent(PGSK buffer: 0.52 g/l Na₂HPO₄.H₂O, 8.8 g/l NaH₂PO₄. 2H₂O, 2.8 g/l NaCl,0.372 g/l KCl, 11 g/l glucose) with glass beads. The protein extractisolated from this was separated by means of isoelectric focusing andthereafter by means of SDS PAGE. The gels were dyed with silver (cf.FIG. 2a) or Coomassie (cf. FIG. 2b). The protein spots, which werevisible only in one of the two gels were cut out of the Coomassie-dyedgels and their sequence determined. It could be shown that theidentified proteins were formed only in Sc5314 in α-MEM medium (FIG. 2aand 3).

[0090] Based on the amino sequence, which was unambiguously determinedby Edmann decomposition of tryptic fragments of the protein, thecorresponding DNA sequence could be identified via data basecomparisons. The DNA sequences as well as the flanking regions wereamplified and cloned by PCR from genomic DNA of Sc5314. Furthermore, thecorresponding DNA sequence from genomic libraries (Liu et al., 1995,Science, 266, 1723-1726) was isolated by means of hybridization of theradioactively marked fragments obtained by PCR.

[0091] The coding sequences for each of the seven identified proteinswere removed from the cloned PCR fragments, by means of PCR, andreplaced by selection markers (URA3) (Fonzi and Irwin, 1993, Genetics,134, 717-728). These constructs are used for the deletion of the codingsequence in C. albicans. Furthermore, the open reading frame for allseven proteins as well as the termination sequences were isolated bymeans of PCR and cloned in vectors with regulable PCK1 and MET3promoters (Leuker et al., Gene, 1997, 19, 192(2), 235-40; Care et al.,Mol. Microbiol., 1999, 34(4), 792-8.) for the expression in C. albicansor with regulable GAL1-10 and MET25 promoters (Mumberg et al., NucleicAcids Res. 1994, 25, 22 (25), 5767-8.) for the expression in S.cerevisiae and in suitable vectors (pMAL, PGEx, etc.) for expression ofthe proteins in bacteria.

EXAMPLE 2

[0092] Detection of the Hyphen-Specific Expression of the ProteinsAccording to the Invention

[0093] The regulation of the hyphen-specifically expressed proteinstakes place at the transcription level since the mRNA for all sevenproteins can be detected only in Sc5314 cultures washed in α-MEM, not inSc5314, which was cultivated in complete medium or in the non-virulentstrain Can34 (Δcph1Δefg1) that was cultivated in complete medium orα-MEM medium.

[0094]FIG. 2b shows as an example a Northern analysis of RNA from thestrains Sc5314 and Can34 (Δcph1Δefg1) cultivated in complete medium. Inthis Northern analysis moreover RNA of the strains Can16 (Δcph1)cultivated in α-MEM medium and Can33 (Δefg1) (Lo et al., 1997, Cell, 90,939-949) were plotted. Can 16 (Δcph1) was described as a strain, whichhas a virulence comparable to Sc5314 and shows the formation of hyphae.Can33 (Δefg1) shows on the contrary no formation of hyphae and itsvirulence is sharply reduced (Lo et al., 1997, Cell, 90, 939-949).

[0095] In the cultivated strain Sc5314, which was cultivated in α-MEMmedium, the corresponding mRNAs can be detected with Cap33-specific,Cap18-specific, and Cap19-specific probes. On the contrary, nocorresponding mRNAs can be detected with the above-mentioned probes inSc5314 that was cultivated in YPD medium. In the virulent strain Can16cultivated in α-MEM medium the corresponding mRNAs can also be detectedwith the use of the probes. The non-virulent strain Can34 (Δcph1Δefg1)cultivated either in YPD medium or α-MEM medium contains nocorresponding mRNAs. The strain Can33 cultivated in α-MEM medium alsocontains no corresponding mRNAs.

[0096] The results of these Northern blot analyses show on the one handthe hyphen-specific expression of the proteins with SEQ ID No. 5, 5[sic], 7, 8, 14, 16, and 18 in the virulent strains Sc5314 and Can16. Onthe other hand these results show that the hyphen-specific expression ofthese proteins is regulated principally at the transcription level. Thisis significant for use as diagnostic means since the presence, in asample to be tested, of mRNA, which is associated with one of theaforementioned hyphen-specific proteins points toward the occurrence ofhyphally growing virulent forms of Candida in this sample.

[0097] Protein extracts from the strains Sc5314 and Can34 (Δcph1Δefg1),which were cultivated in α-MEM medium, were accordingly subjected to a2D gel electrophoresis according to customary processes. As is to beseen in FIG. 3 the hyphen-specifically expressed proteins p15, p18, p19,p20, p33a, p33b, and p40 can only be detected in the hyphally growingstrain Sc5314 but not in the non-virulent strain Can34 (Δcph1Δefg1). InFIG. 2a, moreover it is shown that the strain Sc5314 cultivated in α-MEMmedium produces the proteins Cap33a and Cap33b, the strain Can34 doesnot.

EXAMPLE 3

[0098] Detection of the Hyphen-Specific Expression of the ProteinsCoding Nucleotide Sequences by Means of Nucleotide Chips

[0099] On a substrate surface coated with poly-L-lysine, nucleotidesequences are fixed, which code the hyphen-specific proteins CAP33,CAP19, and CAP18. The nucleotide chip thus obtained was hybridized undercustomary hybridization conditions with cDNAs which were produced frommRNA of the hyphally growing strain Sc5315 and the non-virulent strainCan34 (Δcph1Δefg1) growing in a yeast-like manner. The results of thishybridization are presented in FIG. 1. While the cDNA from thenon-virulent strain hybridized with none of the nucleotide sequencescontained on the nucleotide chip, the cDNA of the virulent strain showedstrong hybridization signals with all three immobilized nucleotidesequences. This experiment shows that with the use of a nucleotide chipaccording to the invention specific nucleic acids can be detected andthat thereby hyphally growing virulent C. albicans strains can bedistinguished from non-hyphally growing non-virulent C. albicansstrains.

1 18 1 900 DNA Candida albicans 1 atgtctaaag tctcaattac tatcatcggtttgaatggtt tcttaggtaa accagttctt 60 gaagctatca attctggtat ttttgatgataagatcaact tcccaatcaa ggcaattacc 120 agaaaggaac cagaaactaa gaatgacaaaattgaatatg ttgtttctga aatcaatgaa 180 gaatcaatta aatcaacttt gagccaaaaattatctggta ctgatgttat tattgaatta 240 attggtccaa atccagaggc tttcgccaatatcgaaaatt tagttgatgc aattaaacca 300 aaattattta tcccatcaca atttggtactgatattccta aagttgatga atatgctcca 360 gggtttttag gaattaaaac tcaacattcagaaaatgtca gaaaatcagg agttaaagtt 420 gttgatatta taacttcgtt atttgctgttccaggagctt ttctttatga atgggttggt 480 tcaactggta ttaatgctga agacagaactgttaaactca ttggtgacat taatcaacaa 540 tttgatattt ctaaattaga agatgttggtaaagctgtac tttctattgc tactaatcct 600 aatccaagag aattaccaga taccattagaattggttctg atagaattac tgttaaagat 660 gtaattgata gatactctaa agatcataatgttgaattga aaattgtttc agaacaatct 720 gcagaagacg ccaagaaaga gtttactgaatctttgaaag ttggttttga tggtgataaa 780 ttcttatggt atttacaagt tattgctgctcaaggtttag ataaaggttt actctccagt 840 aaattggata atgaattggt taatccaggtgaatctttat ggaaatgggg caagtactaa 900 2 900 DNA Candida albicans 2atgtctaaag tctcaattac tatcatcggt ttgaatggtt tcttaggtaa accagttctt 60gaagctatca attctggtat ttttgacgat aaaatcaatt tcccaattaa agctattaca 120agaaaagaac cggaaactaa aaatgacaaa attgaatacg ttgtttctga aatcaatgaa 180gaatcaatta aatcaacctt gagccaaaaa ttatccggta ctgatgttat tattgaatta 240attggtccaa atccagaggc tttcgctaat atcgaaaaat taattgatgc aattaaacca 300aaattattca ttccatcaca atttggtact gatattccta aagttgatga atatgctcca 360gggtttttag gaatcaaaac tcaacattca gaaaatgtca gaaaattagg agttaaagtt 420gttgatatta taacttcgtt atttgctgtt ccaggagctt ttctttatga atgggttggt 480tcaactggta tcaatgctga tgacaaaact gttaaactta ttggtgacat taatcaacaa 540tttgatattt ctaaattaga agatgttggt aaagctgtac tttctattgc tactaatcct 600aatccaagag aattaccaga taccattaga attggttctg atagaattac tgtcaaagat 660gtcattgata gatattctaa agatcataat gttgaattga aagttgtttc tgaacaatct 720gcagaagatg ccaagaaaga gtttactgaa tctttgaaag ctggttttga tggtgagaaa 780ttcttatggt atttacaagt tattgctgct caaggtttag ataaaggttt actctccagt 840aaattggaca acgaattggt caacccaggt gagtctttat ggaaatgggg caagtactaa 900 3501 DNA Candida albicans 3 atggcctcct cagtaaagtt ggctacggca cttaaacaacgtgctatatt gacaaaagaa 60 ttgtctgaat tagatgataa aatacaatct tcattgattctgcaagttgg tatgaaaaaa 120 atcaatgatc cagataaatt gtatttagat tatgttgctaaatctcaaga attggctaaa 180 ttggtatcat caataaatta tactaataat ataactccaattgaacttga tttgacaatg 240 ggaaagtatg ataatactat aaaaacaatt aatgatgcattaatttgtcg agaccgaata 300 tttaaaaaat tacaatttgt gaaaaaaata tcaacagcaggtaaagaaca accattagat 360 tccaaagatg aaattaaatt tgtatcattt attgatgttgataaatatga tactttggcc 420 caagaattaa atactcaatt tgagaatttg aatttgaaattacaagaaat aaattggcaa 480 gttgatcttg ttgagatata a 501 4 486 DNA Candidaalbicans 4 atgaaattag ctgaagcatt aaatttaaaa aagaacttgg aaagagatgctggtgaactt 60 aaatcattaa ttcttaaatg ttgtcaagct caaactggcg aaaaccctccatttgatcct 120 aatgaattat ttgaacaata tgaagaaatt gataaattaa ttactgatataactattaaa 180 atacaacgaa ccaataatga aataaagttt gcctatgata atgataataagtctaatgaa 240 gaaccacttc gatcaatgac acaagctatt gctgatattg atgatttagaaagacaaatc 300 aatgtgacag atgatataat tcataatggt attattacaa aactgtattcgaccaagaag 360 attgctgatg tgtcacatgt tgacgtggtt gcatatgaca agacaagaaagaaaatgaat 420 gagagattag ataaattaaa acttcgtata cagtcggcaa attgggaatttgatctaatt 480 gattaa 486 5 299 PRT Candida albicans 5 Met Ser Lys ValSer Ile Thr Ile Ile Gly Leu Asn Gly Phe Leu Gly 1 5 10 15 Lys Pro ValLeu Glu Ala Ile Asn Ser Gly Ile Phe Asp Asp Lys Ile 20 25 30 Asn Phe ProIle Lys Ala Ile Thr Arg Lys Glu Pro Glu Thr Lys Asn 35 40 45 Asp Lys IleGlu Tyr Val Val Ser Glu Ile Asn Glu Glu Ser Ile Lys 50 55 60 Ser Thr LeuSer Gln Lys Leu Ser Gly Thr Asp Val Ile Ile Glu Leu 65 70 75 80 Ile GlyPro Asn Pro Glu Ala Phe Ala Asn Ile Glu Asn Leu Val Asp 85 90 95 Ala IleLys Pro Lys Leu Phe Ile Pro Ser Gln Phe Gly Thr Asp Ile 100 105 110 ProLys Val Asp Glu Tyr Ala Pro Gly Phe Leu Gly Ile Lys Thr Gln 115 120 125His Ser Glu Asn Val Arg Lys Ser Gly Val Lys Val Val Asp Ile Ile 130 135140 Thr Ser Leu Phe Ala Val Pro Gly Ala Phe Leu Tyr Glu Trp Val Gly 145150 155 160 Ser Thr Gly Ile Asn Ala Glu Asp Arg Thr Val Lys Leu Ile GlyAsp 165 170 175 Ile Asn Gln Gln Phe Asp Ile Ser Lys Leu Glu Asp Val GlyLys Ala 180 185 190 Val Leu Ser Ile Ala Thr Asn Pro Asn Pro Arg Glu LeuPro Asp Thr 195 200 205 Ile Arg Ile Gly Ser Asp Arg Ile Thr Val Lys AspVal Ile Asp Arg 210 215 220 Tyr Ser Lys Asp His Asn Val Glu Leu Lys IleVal Ser Glu Gln Ser 225 230 235 240 Ala Glu Asp Ala Lys Lys Glu Phe ThrGlu Ser Leu Lys Val Gly Phe 245 250 255 Asp Gly Asp Lys Phe Leu Trp TyrLeu Gln Val Ile Ala Ala Gln Gly 260 265 270 Leu Asp Lys Gly Leu Leu SerSer Lys Leu Asp Asn Glu Leu Val Asn 275 280 285 Pro Gly Glu Ser Leu TrpLys Trp Gly Lys Tyr 290 295 6 299 PRT Candida albicans 6 Met Ser Lys ValSer Ile Thr Ile Ile Gly Leu Asn Gly Phe Leu Gly 1 5 10 15 Lys Pro ValLeu Glu Ala Ile Asn Ser Gly Ile Phe Asp Asp Lys Ile 20 25 30 Asn Phe ProIle Lys Ala Ile Thr Arg Lys Glu Pro Glu Thr Lys Asn 35 40 45 Asp Lys IleGlu Tyr Val Val Ser Glu Ile Asn Glu Glu Ser Ile Lys 50 55 60 Ser Thr LeuSer Gln Lys Leu Ser Gly Thr Asp Val Ile Ile Glu Leu 65 70 75 80 Ile GlyPro Asn Pro Glu Ala Phe Ala Asn Ile Glu Lys Leu Ile Asp 85 90 95 Ala IleLys Pro Lys Leu Phe Ile Pro Ser Gln Phe Gly Thr Asp Ile 100 105 110 ProLys Val Asp Glu Tyr Ala Pro Gly Phe Leu Gly Ile Lys Thr Gln 115 120 125His Ser Glu Asn Val Arg Lys Leu Gly Val Lys Val Val Asp Ile Ile 130 135140 Thr Ser Leu Phe Ala Val Pro Gly Ala Phe Leu Tyr Glu Trp Val Gly 145150 155 160 Ser Thr Gly Ile Asn Ala Asp Asp Lys Thr Val Lys Leu Ile GlyAsp 165 170 175 Ile Asn Gln Gln Phe Asp Ile Ser Lys Leu Glu Asp Val GlyLys Ala 180 185 190 Val Leu Ser Ile Ala Thr Asn Pro Asn Pro Arg Glu LeuPro Asp Thr 195 200 205 Ile Arg Ile Gly Ser Asp Arg Ile Thr Val Lys AspVal Ile Asp Arg 210 215 220 Tyr Ser Lys Asp His Asn Val Glu Leu Lys ValVal Ser Glu Gln Ser 225 230 235 240 Ala Glu Asp Ala Lys Lys Glu Phe ThrGlu Ser Leu Lys Ala Gly Phe 245 250 255 Asp Gly Glu Lys Phe Leu Trp TyrLeu Gln Val Ile Ala Ala Gln Gly 260 265 270 Leu Asp Lys Gly Leu Leu SerSer Lys Leu Asp Asn Glu Leu Val Asn 275 280 285 Pro Gly Glu Ser Leu TrpLys Trp Gly Lys Tyr 290 295 7 166 PRT Candida albicans 7 Met Ala Ser SerVal Lys Leu Ala Thr Ala Leu Lys Gln Arg Ala Ile 1 5 10 15 Leu Thr LysGlu Leu Ser Glu Leu Asp Asp Lys Ile Gln Ser Ser Leu 20 25 30 Ile Ser GlnVal Gly Met Lys Lys Ile Asn Asp Pro Asp Lys Leu Tyr 35 40 45 Leu Asp TyrVal Ala Lys Ser Gln Glu Leu Ala Lys Leu Val Ser Ser 50 55 60 Ile Asn TyrThr Asn Asn Ile Thr Pro Ile Glu Leu Asp Leu Thr Met 65 70 75 80 Gly LysTyr Asp Asn Thr Ile Lys Thr Ile Asn Asp Ala Leu Ile Cys 85 90 95 Arg AspArg Ile Phe Lys Lys Leu Gln Phe Val Lys Lys Ile Ser Thr 100 105 110 AlaGly Lys Glu Gln Pro Leu Asp Ser Lys Asp Glu Ile Lys Phe Val 115 120 125Ser Phe Ile Asp Val Asp Lys Tyr Asp Thr Leu Ala Gln Glu Leu Asn 130 135140 Thr Gln Phe Glu Asn Leu Asn Leu Lys Leu Gln Glu Ile Asn Trp Gln 145150 155 160 Val Asp Leu Val Glu Ile 165 8 161 PRT Candida albicans 8 MetLys Leu Ala Glu Ala Leu Asn Leu Lys Lys Asn Leu Glu Arg Asp 1 5 10 15Ala Gly Glu Leu Lys Ser Leu Ile Leu Lys Cys Cys Gln Ala Gln Thr 20 25 30Gly Glu Asn Pro Pro Phe Asp Pro Asn Glu Leu Phe Glu Gln Tyr Glu 35 40 45Glu Ile Asp Lys Leu Ile Thr Asp Ile Thr Ile Lys Ile Gln Arg Thr 50 55 60Asn Asn Glu Ile Lys Phe Ala Tyr Asp Asn Asp Asn Lys Ser Asn Glu 65 70 7580 Glu Pro Leu Arg Ser Met Thr Gln Ala Ile Ala Asp Ile Asp Asp Leu 85 9095 Glu Arg Gln Ile Asn Val Thr Asp Asp Ile Ile His Asn Gly Ile Ile 100105 110 Thr Lys Ser Tyr Ser Thr Lys Lys Ile Ala Asp Val Ser His Val Asp115 120 125 Val Val Ala Tyr Asp Lys Thr Arg Lys Lys Met Asn Glu Arg LeuAsp 130 135 140 Lys Leu Lys Leu Arg Ile Gln Ser Ala Asn Trp Glu Phe AspLeu Ile 145 150 155 160 Asp 9 6619 DNA Candida albicans 9 gttgataagatctctaatga tgatttgtaa tttgagcgaa tttttatctc ttgttgggtt 60 tttgtggatgttgcacataa agctgcaagg acatcaccaa caacaagtag caagtgtggc 120 tagagttacaaatccgtgta tggtagcaca actgatgaca tttgaataga tgtcatacaa 180 caaaatatggaatagttttg gataataaac agcacgtgac tattgttaac cagatggctg 240 ttgagaagacactaagacag tacaacagat atctacaaac acctataggt aaatgaggac 300 tgcctatttccttgaaacca ttttctatta cttatttaca ttagttgtat cttttcatta 360 attcaatttcattcataaat atcaaatacc tagtatctaa ctacatattg cctactttaa 420 atgaaaataaaccttgggac ataataaatt atatctgata atcttagtac ttgccccatt 480 tccataaagattcacctgga ttaaccaatt cattatccaa tttactggag agtaaacctt 540 tatctaaaccttgagcagca ataacttgta aataccataa gaatttatca ccatcaaaac 600 caactttcaaagattcagta aactctttct tggcgtcttc tgcagattgt tctgaaacaa 660 ttttcaattcaacattatga tctttagagt atctatcaat tacatcttta acagtaattc 720 tatcagaaccaattctaatg gtatctggta attctcttgg attaggatta gtagcaatag 780 aaagtacagctttaccaaca tcttctaatt tagaaatatc aaattgttga ttaatgtcac 840 caatgagtttaacagttctg tcttcagcat taataccagt tgaaccaacc cattcataaa 900 gaaaagctcctggaacagca aataacgaag ttataatatc aacaacttta actcctgatt 960 ttctgacattttctgaatgt tgagttttaa ttcctaaaaa ccctggagca tattcatcaa 1020 ctttaggaatatcagtacca aattgtgatg ggataaataa ttttggttta attgcatcaa 1080 ctaaattttcgatattggcg aaagcctctg gatttggacc aattaattca ataataacat 1140 cagtaccagataatttttgg ctcaaagttg atttaattga ttcttcattg atttcagaaa 1200 caacatattcaattttgtca ttcttagttt ctggttcctt tctggtaatt gccttgattg 1260 ggaagttgatcttatcatca aaaataccag aattgatagc ttcaagaact ggtttaccta 1320 agaaaccattcaaaccgatg atagtaattg agactttaga cattgtgata gatagatatt 1380 atagattaattattagataa gcttgtgtaa ttgatcaatt gcttgattaa tgagattgga 1440 aaacaaaaaattacaagcca tgttgaatgg aggaaacacg tctatttata atgggtttga 1500 ttcaatgtgatgcttaatag gggagtgggg ggttatgcaa tgtaaggaga gacgacaaaa 1560 catacttagctaaaaacaca aacacacatt gttgccatag ttaaatgtgg aattaaatgg 1620 aacaatcttttcccgtaaaa tgtaaagaaa ggaggaaaaa catacaccaa gaaattgtgg 1680 cgtaatctgaaattctttgt ttctctcttt ctctgtttaa tttgtaatca aatatttttc 1740 tcattacataatatgcaagt gatgattaat aatcaatatt tgtttatcag ttatatctat 1800 ttaatccttgtatttataat ttcataacaa atcaataaca acacccgcta cagccacatc 1860 acaatcaatttactggtaac ttatttgtaa tctacatatt acctaagatt gtacagaaat 1920 tgtttctgcttactaaattg tattggtaat aattctacta tggagtaaat agtgttgcta 1980 ttataattgtggctagtgta tatactgata tcaattaact cgtattaata atatattagg 2040 tgtagcaagcttgatatctt tgacacagct gttatttgtc tacgccacat tagatttctt 2100 aaccacaacttgtaaggtag tatagacaac taaaacccta tacagggctc tcttaatcct 2160 tcctccaatgtctgactact aagtaccatc ataccaccag ggttaggtag atatatagac 2220 tctcttggatgcccttgtcg tctactccta aagaggttta catactcaac tagaaaatac 2280 aagctattaacgtagttaaa gtattacaca actttgtata gctgactttg caaccctcag 2340 attcgttgaattttttttgt aagtaacaat ctgtggtctc tggtaaatag ctcaagcctc 2400 agttattactaatcttatta cttatactgc caattctaaa accatcagca aattcattat 2460 taagcattggttttagattt catttggtta gtaaagttgc agatacgata tacataacta 2520 agcatagactataataatcc gaaactaaaa gttgcaatat tccgatatcc ggtatccttt 2580 ccgttgaatcttttgtattc taaatattaa tacaaggtat agcttgatta attgttggtt 2640 tatgtagatgacagtacttt acttattggt tgtaaataat ctaaagaagt acagaattta 2700 tttcaaaattggtatccaaa ctcaaccaaa ataataaatc aaccacccaa attgatatta 2760 tctaaaactacaaataaaga tatacaaatg gtttagccta ctgaaagtat acataatagc 2820 atcagttaactcctcgtagt tcaaccaact aaaaataacc tttataatgg ttctcatctc 2880 tagttgtatcattctgttcc tcatcctcgt catcttcatc ttcgtttgaa ttatcttcaa 2940 cagtatataacccaattctt cttcgattat tattattgtt atttgatgat agctgtttat 3000 tctttttaaatttcttgaaa tttttccttc cagcatatct cgattcagta tcatcatcaa 3060 cactcttatttttgttacgt cgatttctag gattaactaa atcaacaatt tccacaattg 3120 ccaaattttgtaaattttca ttaatttcaa attcagattc caaatctaaa tttgatgatt 3180 tattcaattcttgaactttt aaaactgcat ctactaatga tacttttggt gtaaattttg 3240 gaattttaaatggtttttga tcatcattag tagtttcatc tttattctta cgttttgatc 3300 cgattgcaggaatttcttcc tcgtcttctt gttcttcttc ttcattttta attgatggaa 3360 aattggaactagacttttgt gaattggaat ctggtttaga tttccttgct aattgatcag 3420 ttgatgatgataaattcgaa tcacccaaac tattaagtgc atcttcttgg ttgattttac 3480 gactttgcgtatgagaattc aacacttttt gattttgctt tatcgttgaa atttttttaa 3540 tgggatcacttaattgtgtt atttcttgag ttattgaatc attatcacca ctggtattaa 3600 ttttctgactatttaattct ggtgatgtgc ttttattgtc agtgtccaag attgatactt 3660 taggggatggatcacgtgta gatggtactg gtgtagttgt agttgttggt gttgtcgtac 3720 ctaaggagaagaaatgtaat ctatcaactt tttcatattt ccttctcttt ctccctgaag 3780 ggtgccgccacctcaggtgt ttgtgttgta actgatttat catctgatgc taacatagtt 3840 gttgaggtacgtttcaatta aatcaaataa tttttgggaa gatctagtat tgagccttct 3900 ttaattgattgccataattg atcttctgtt agagtttgga aattatgtct attcaaagtt 3960 gttgccattgtagttggaac aattaaaatg aattgattat cacctatttt cgatttcaat 4020 acgtcttcttgttgttgtat taacacttta aaatctataa ttattgtttc caaccaattt 4080 attctctccaatgatactaa tttaacatta tttaatggct ttaatctata acttttcggt 4140 aaatagtgatcatccttttt tggtaagtat ttcttataat ctatatccaa aagccaatcc 4200 ttaacactattatacccatc ctcattatca tgacctaata ctttttgtaa ccattgtgaa 4260 tttaaaattgggataccctt aataattgcc aatttgaatt catctttttg ttcttcctcc 4320 ttgatatcgttattatcact atcataataa taatgagtgg catccattat attatcaacg 4380 atttgaatatcaataccatg atcttttatt attgttttgt aatgattttg atcacatttt 4440 tcattcgcactaatatataa tttaaaggga atccattcaa tgaacaatgg agtatcttga 4500 gttattatttgtcctgtttg tttatcaatt tgtggaatag tttcaatttt caatttgaat 4560 tcatttgtgaaatcaatttc ttctggttta ctatcttttg ataataattt atatgttttc 4620 ccatttataatcgttttcgc tcgactacat attctaatga ttaataatgt tttatcacta 4680 cctattgaagaagaagacga tgacgatgat ggtggttggg tattgcctat gttgggtcca 4740 gttattaattcaatcaattc tcttgatact ttaggtgacc agaattttat atcagtattt 4800 tcatcacgtcccacggtata atttgtttct ggtgataaat gtttatattc taatgaattg 4860 ttagttggcctggggttaaa taaatatcaa aaatcttcag tttctaacat acctcctctt 4920 ggatcatctttatgatatgt tatccacata gttaagtgta catttgcttg tcaatgaagt 4980 cttcctaagcgaatttgaac tgaaaaaaaa cctatcgcga tccctttaca actttaacaa 5040 acgtataataaacaggttgc ataaaacctt ctacaaacca ttcagaatct tactttcata 5100 aatagtgatttattatgaat cgtttcttta aactagtaat acttgtatca ataatgaatc 5160 cctaagcttttatgttgtaa ctggattaat atcagagttg taggcgtggt cacgtgacat 5220 agatagaataagagtcgaag ggaacaacat taattagact tgatacttat tgtattaaga 5280 taaatgtgaatttacaataa caaatggtga atatagttta tcagttcata ctagacgatt 5340 agttgaaatttatttatgtt tgtttggata atcgtttgtt ataaaaatag aaatcgtcat 5400 tttttttttgctcacttgtc ctgttcattc cattcaaggc tgtaactgta aatgttaaac 5460 taaatttcatttcatttcat tcaacaaaaa aatatcatac tctatttaac ttcaacaaac 5520 taatctcaagaaccagctta cttcctttta taatactaca acaacatata ttatacaact 5580 aatcaaagatgggtattaag aaaatgtttc aaaagaaaga accaaccgaa caagaaattc 5640 gtgaagaattaagtcgagtt ggcatttcta caagatcaaa taatactcga caagagaagt 5700 ttggtgcatttaaaaattat gctcaagaac gagctaatat gaaaccacaa ttaggaccag 5760 ttggtggtaatccttatgcc aatattaatc ctgggaccaa caataataat aataatccat 5820 atgccaatgataatgggaat aatagtactg gcaaccccaa caacaacaac ggtggtaacc 5880 cctatggtggtggtgttact aataataatc cttatggagg ctctggtggt aatggaagag 5940 gatcatcacctagtccttat gcaccgacta catcaacaac tactagatca tctaatccat 6000 atggaaacaataatggttct agatcaagtc aaaacacttc tagtccttat gccaaatcaa 6060 ctaacaattcatcatattct aactcaccgt attctggatc aactgtaaat aatggtaatc 6120 gtggcggccatagcaacaac agcaatagtt ctgctggtgg taacccttat gctgccggtg 6180 gtagaagttcacaatctcaa aattcacgag acaatgtata tacagctcct gccactcgta 6240 catcaactagacaaactcaa ggatatggag gtggtgatac cgattcgact cttgacctta 6300 atgccattccatcacatcaa atgtttgata ataagaaacc gatcaaaaga aatcaacaaa 6360 gttcacaacaacctgccaat gattataatt tagatttaaa tgatgaatat ggcgaagaag 6420 aagacttgaatttggatata agtgaagtac ctgaagaaca acaacaaatc aattctgaag 6480 atgaagaagtagaagccatt aaacaagata ttaaatttgt caaacaagaa tcagttcaaa 6540 gtaccagaaatactcttaga atggcacaag aagctgatgc atcgggtact aatactttag 6600 gaatgttaggatcgcaact 6619 10 7965 DNA Candida albicans 10 tagctaagta tgctttgtcgtctctcctta cactgcttaa ccctccaccc ctctatttgg 60 cttcacattg aatcaaaccattataaatag acgtgtttcc tccattccac atgacttgta 120 attctttgtt ctccaatctcatcaatcaat tacacaagct tatctaataa ttaatctata 180 atatctatct atcacaatgtctaaagtctc aattactatc atcggtttga atggtttctt 240 aggtaaacca gttcttgaagctatcaattc tggtattttt gacgataaaa tcaatttccc 300 aattaaagct attacaagaaaagaaccgga aactaaaaat gacaaaattg aatacgttgt 360 ttctgaaatc aatgaagaatcaattaaatc aaccttgagc caaaaattat ccggtactga 420 tgttattatt gaattaattggtccaaatcc agaggctttc gctaatatcg aaaaattaat 480 tgatgcaatt aaaccaaaattattcattcc atcacaattt ggtactgata ttcctaaagt 540 tgatgaatat gctccagggtttttaggaat caaaactcaa cattcagaaa atgtcagaaa 600 attaggagtt aaagttgttgatattataac ttcgttattt gctgttccag gagcttttct 660 ttatgaatgg gttggttcaactggtatcaa tgctgatgac aaaactgtta aacttattgg 720 tgacattaat caacaatttgatatttctaa attagaagat gttggtaaag ctgtactttc 780 tattgctact aatcctaatccaagagaatt accagatacc attagaattg gttctgatag 840 aattactgtc aaagatgtcattgatagata ttctaaagat cataatgttg aattgaaagt 900 tgtttctgaa caatctgcagaagatgccaa gaaagagttt actgaatctt tgaaagctgg 960 ttttgatggt gagaaattcttatggtattt acaagttatt gctgctcaag gtttagataa 1020 aggtttactc tccagtaaattggacaacga attggtcaac ccaggtgagt ctttatggaa 1080 atggggcaag tactaagattatcagatata atttattatg tcccaaggtt tattttcatt 1140 taaagtaggc aatatgtagttagatactaa gtatttgtta ttaataaatc agattaaatc 1200 aatgaaagga ttacaattaatgtagatgag gattagaaat tggtttctaa tagagaactg 1260 ttgtactctc ttggtgccttctcaacagcc atctggttaa cagtagtcac gtgctgttga 1320 tcatccaaaa ctattccacattttgcttta tgaaaaccta ccctaatatt gccaattgtg 1380 ctaccataca cggacttgtaattccagcca cactttctta cttgttattg ttgatttcct 1440 tgcagcttta tttgcaacatccacaaaaac ccaacaagac aaaaataacc tcaaattaca 1500 aatcatcatt aaagatcttatcaacaacat gtcccctttt tgataattaa caccttaaat 1560 taacctcttg tggggttgatcaaaccttga atttgaaccc taagtgcaag taagaacgaa 1620 gttaaagcat ccaagtcttgtgaaaaagca acatgtaacg gcattttagt ttcgaaccga 1680 gttaaaaatt ggaagtaagaataagagaaa ctgcgtgacg gctgtgcggc atattattta 1740 aggcaaattg acgatggcaaaaaaaatctt taatttttcc gtcttggtaa ttaaatatcg 1800 cgaaggcatt cctttaaagtaatgccttta tgaatggtca aaatgcccta ttttaaaatg 1860 ccttattact ttctattgcgcttatcatga aatttggcta ttactggcac ggcaacaatt 1920 taatcagagc atctgaaatgtgggtgatta attgcagagg gcggttagat ataattttag 1980 atggattgat ttgcctaattaggaatttgt aactttataa acctctttat aatttttcat 2040 cttttcattt ttttcaaaataaaatgggtt tttcacagaa gtaaagatat tgttcatgct 2100 tattgtgcta acctgatgtaaatctcttta ttttaatttg acttcttctt caaaccgttg 2160 aacacggcac caatgaatgataaaattgat gatatttagt taccagcaag caatgatgac 2220 agtattccca agaagaagagacaatggcta cagcaactgg agtaaaacaa gtagtttagc 2280 tacagatcta acctttttgtaacaccccaa ccattctctc tttttttgct atattgaaac 2340 actgattttt cttaaattatttacccaact cctaatggaa tataacaacg aggtgcttct 2400 acttctccac atttccaacaataaaacatg aatcaacatc aatattgttg caattaatca 2460 aactcttaac atctccacctccccactaat gatcgatatt atatcaaaac cattggaaat 2520 ttagtttggg tttcatttgaatttcggtca agaaaattaa aagtaaaaaa gaaaaaaaaa 2580 atttattatt attattcggttcgatattgc cgcaaaacca aattccatca tcatttcaca 2640 atataatata aaaagtcttcaatcttacac cttgcaaaaa gtttcaattt ttttttataa 2700 aatatttatc tatattctaattgttacatt tattctttac ttctaatcaa aacaactata 2760 tatcaatatt atgtttgaagtaggtgaaaa atatcctgtt gaaagcagca gtagttcaaa 2820 tgacatagaa tctcgtggtgttcaacctat aacatccctc aaagacaata aatcaatagg 2880 aatgatagag aaagataatgatgatctatc atgtgaacaa tatagtactt gtgatgaagt 2940 caaaagagat ttaaaagcaagacatgtttc tatgattgcc attggtggta caataggtac 3000 agggttattc atatccactggttctttact tcacaccact ggtccagtaa tgtcattaat 3060 atcattttta tttgtcacaactttagcata ttcagttaca caatcacttg gggaaatgac 3120 aacatatatc cccgtttctggatcatttgc ccaatttata actcgttggg tttcaaaaag 3180 ttgtggtgct gctaatggttggttatattg gttttcatgg gccataacat tcgctttaga 3240 attatcagtt gttggtcaagtcatacaata ttggactgat gctgtaccat tagctggttg 3300 gatttccatt tttttcgtcttattaactac atttaattta ttcccggtga aatattatgg 3360 agaggttgaa ttttggattgcttcaactaa agtaattgct attgttgggt ggctcatata 3420 tgcattttgt atggtttgtggggctggtaa aactggacca gttgggttcc gttattggcg 3480 gaatggatat gcatggggtgatgggatgat agttctgaat aatggtaaat acgccatttc 3540 tttcattaat ggtcttatcaatgctgtttt tactttccaa ggtactgaat tggttgctgt 3600 tactgcgggt gaagcttctccaagagcaat ccgtagtgca attaaaaaag tcatgttcag 3660 aattttggta ttctatgtcttgtgtatgct tttcattggt cttttggttc cttacaacga 3720 tccaaagctt actcaagatggtggttttac aagaaactct ccattcctta ttgctatgga 3780 aaattccggt actaaagttttaccacacat tttcaatgca gtgatcgtta caacaattat 3840 ttcagctggt aattccaatgtttattcggg atcacgtatt ctttacgggt tagcccaagc 3900 tggtgtagct cctaaatttttccttaaaac taacaaaggt ggtgttccat attttgctgt 3960 cttgttcact gcggcatttggtgcattggg atatttagca tgttcggaag atggtaataa 4020 agctttcact tggttattgaatattattgc cactgctgga ttgatcgctt ggggattcat 4080 ttctgtgagt catgtcagattcatgaatgt tcttagaaaa agaggtttaa gtcgagacat 4140 tttaccttat aaagcttttttcatgccata tagtgcatat tatgccatta ttattatatt 4200 cattgttgtg ttgattcaaggtttcacagt gttttgggac ttcaatgcta gtgatttctt 4260 cactgcctat atatccgtgatattatttgt tgttctttgg attggtttcc actttttctt 4320 ttacgggttt ggtaaagattcttttaaatg ggaaaacata ttaatcccat tggatgattg 4380 tgatattgat tctggtgttagagatattaa tgatgctgaa tttgatgtac ctgaacctaa 4440 aaatgtttgg gaaagattctggttacttat tgcttaatct taatttatat aatttaatac 4500 ttaatggaca tagagcttttcagcgattat aatagaaact gattatactt atttaattta 4560 aatctattta caaatttctctgaagagtgg ggcgccggtt atacaaactc aacaacgtaa 4620 taattctgtt taccaactctaaccaacaac aatttaccat caatcaaatg attatcaaca 4680 tcaaataata taacatcatctggatcctca atttgatttc tgtccaatcc catgtataca 4740 ccaccagctt taatcaatcttctcatttca ccctttgatt taccaacaat atcagctaat 4800 aatgcactca atttgatttcttcatcaggt gaaggcttat ttcttttaaa caaaatacct 4860 gatcttttaa aattttctatcaatttatca gcgcttacat tatcattaaa tggttgatct 4920 ggagtaggga ataaaaatcccgtaataaac gccatttcgt caccaacacc aacaccatgg 4980 atcaaatcaa caacttcacgtgctaaaaca cgttgagcaa tacgtaaacc aggatcactg 5040 ttatgtttag gtaataattcaccttcaatt acattcaagg gcaataatgt gaacactttt 5100 aataatttgc ccactatatcatctggaaca ttaatgaaat attgatacat ttgataagga 5160 gtggtcaaac tagaatcaataaatactgca tttccagcgg atttaccaaa tttctcacca 5220 ctagaagtag tcaataatggaacagtaagt ccataagctt catgtttctt gccatgaaat 5280 ttcttcaaac gtgaaattaaatcaatacca gcagtaatat tcccccattg atcatttcct 5340 ccaacttgca tattaacattttcatccttg tataaatgcc aaaaatcata agcttgtaga 5400 atctgatagg taaattcattgaatccaatt ccacccagtt ctaatcttga ttgaatggaa 5460 tcacgtgcta acatcgaactaactctaata tgtctaccat atgtagccaa aaattccaac 5520 atcttcacgt tttcccaccatgaagcatta tttaccgatg tagtatcacc gactttttca 5580 gtcatgggga attgtcttgatttggcatat tctatcccat tactcaaaaa tgtagaaatt 5640 tgtcgttgga ttttcgtcacattatcttca acttcaactt catcaatctt gttacgctct 5700 gttttcctcc cacttggatctccaacaagt ccagtagcac ctccaacaag tccaacaaca 5760 tcattaccac tcattttgaaatgtaataac accattaatg gtaataaatt acctaaatgt 5820 agtgatgatg ccgtaggatcagcaccacaa tataatttga atttgtgatt agaaccacgt 5880 ttagtcaatt tatataaattatcatcggtt attgattcaa ttaaatgtcg actttgtaaa 5940 tattctaata atgaattgtcgggattggtt tctggtgtta aatctttggc ttcagtcaat 6000 tcatagatgg taggaatgatagtgactgga tctcttgcaa ttgttgaatt aaatctagca 6060 agccttctaa tcaaaggtatgtttctggta tgtgttttca acatattact tgatgtctgg 6120 ttgaacttct ggttgtcgtttcttcgattg aattttttct tgtagcttca ttagcgggct 6180 tatttgctat tcgcggtttaatttttaaag aaagccgcaa attcaaatcc aaatccatct 6240 caagctgaga tttttctttaattttttttt ttttcacttt actgatatca ttctaatcat 6300 taaacataca aagctcctaaaccaatgaca gatcagatta aaggggatga acataaccaa 6360 gtaaatggta gtaaaaaaagaaagagaaag agaaataaga acaagaaaaa tgacaacaac 6420 acaccagtag agacttccgaaccaataccg acacctgttt atgaagatga tatccatcga 6480 caaaataaaa agttcaaattcaatgaggaa ggggaaatgg agaaacctca agagtcacct 6540 gaagaggaac aactagaattagtggcagat caaggggaac cttgcactga agaaccttta 6600 ccacaacatg aaggtttcgaagagattgaa gtaactgacg acatagatga aacagaagaa 6660 ccagagaatc ttccaacaaggactcaacaa gaaaaacatc aacacggtaa gaataaattt 6720 aaacaaaagc ttgaattcaaaagaaaaacc gttgtatata aagatcaaga tgatgaagac 6780 gatgaggaag aaaataatactttcaatttt tcacaaaatt cgtttcaact tgcagccacc 6840 gcccaacaat tgttacaaattagagagaaa ttgcccattt atcatcataa ggataaaatc 6900 attgaatgca ttaataataatcaagtcact atcgtcattg gtgaaaccgg ttcaggtaaa 6960 tcaacacaaa tccctcaatttttaatgcca gaaaacccaa aaatgattgg cgtgacacaa 7020 ccaagaagag ttgccgctgcttctttagca gcaagagtaa gtgaagaata tggatgtaaa 7080 ttaggtcaag atgttgggtatcaagttaga ttcactaata tgactaacag acaaacaaaa 7140 ttgaaatatt taactgatggtatgcttcta cgagaaatca tgcttgatct gaatttgact 7200 aaatattcaa caattatcctagatgaagcg catgaaagaa ctattttgac tgatttaatc 7260 atggggtttt tgaaacaaattattacttct ggtaaaagaa aagatttgaa aatcgtagtt 7320 atgagtgcta ctttgaatgccgaattattt agtaatttct ttgataatgc tcctatttta 7380 tacattgaag gtaaaatgtatccagtttca caattctact tagatgctga atctgaagat 7440 attgtggata ccatgatcagaagtataatt caaatcaatc ttaatgaacc cgagggggat 7500 attctttgct tcttacctgggcaagaggaa attgataatt gtgttaaaag tttagaacaa 7560 ttagcacctc aactacctagggaggcacca ttgattgttc ctttaccttt atatgcagct 7620 ttatcacctg gccaacaatctaaaatattc gaaaaattac ccaagggaag aagaaaagtg 7680 attttggcga caaatattgctgaaacatcc attactgttt ctggtgttaa atatgttata 7740 gattccggat taaggaaaattaaagtttgg aaacataatt taggactttc tacattattg 7800 actaccccta tttcacaagcttcagcaaga caaagagccg ggagagcagg tagagaatct 7860 gaaggtaaag tattcagattatatcctgaa tctacttata tggcacttcc aaaacaacaa 7920 gaatctgaaa ttaaaagaaatgatattatt ttacccagtt ttgac 7965 11 5158 DNA Candida albicans 11aataattatc attagtcaat tcaacaacta taggagattt agcagcaatc tcttgagagt 60tccttcctcg acatctaaca acaacttgga tatttgacat tgatgataat gctgctatga 120gtattaattt aactgaaatc acaagatgaa gaatgaaaac aacaacaaca aagagaaaga 180gtttggcaac gggagaggaa gagaaagtgt aaacaaaaac aaacaaccat aaaaatttac 240accataaaaa aaaattagaa gtcgtgattg aactatatgc aggccactat aagaagatat 300taaaactact ctgattgaat gaatgaatga ttatataaat ccctcttttc tctcaactta 360tagccttaat caaagaaatc atcgtcatct tcatcatcat cagcagctat atttttccta 420ttgaaagtta tttttgtttg ttgcttttgt tgactatctc tacttaatcc atctaatatt 480tgcacaatat ctttttcacc aagtttttga tgtatttgac ccattgaata taatttaata 540atataatttt ctactgcttg agctctatcg ggtctaacaa tctttacacg acttaatctt 600tctctagctt cattagttaa gactcgattt aatatggtta tggtcatatt ctcttgtgcc 660agatcttgtg cgccacccga agaagaagaa gatggattgg tactactgcc acctccggca 720gcatttcttt gtaattctgc taatcttgct tgtcttatag catttaattc tgcgtcatcc 780ataatgtata gttgtgaatg aatggagaag gagtagtatt aaaataatta gtgtggatag 840agtagtcagt cagtcagcca agtgaatagg gaagtaagga aaaattttgg tcacatttaa 900cacgaacttc ttgatcaaag aagaagaaga agaaattttt tttctgtcat cacgtgcacg 960acctttaaat caattgacaa ttcaaaaatt ttgaacaaca acacaacaca actcattctc 1020tctttctctt tctctctctc tctctgtgaa aaaaaaaaaa aaagtaaagg acaataaaga 1080aatcaaacaa tcaattaaac aaagttaaaa caggaacttt ttctattcaa gttcaaattg 1140aaagagaaag agaaatagaa agaaaaaaaa aatttagttc aaattggaat cttgtcttat 1200ttagtttcat ttctatatat cttgtcctca tatactatca acatttagat tgatttgaat 1260ccagaatcaa caatttcaac aattcttcag attttgatat agtgtattct atttgacata 1320ctttactact accaatacag tcacataatt acatatataa atatattaag agtgggtttt 1380cggaacattt tcctcctaga tttaatatag aatccctttc ccctaatttt ttttgcatca 1440acatttactt aaaaacttca accccaccaa ctcctaaccc taatatttcc cctttctttt 1500tttgcatata agactcccac aatgagttca gataaatcaa atttactaaa aaaatacaag 1560attgtctttc ttggtgatca aagtgttggt aaaacatcat taatcaccag atttatgtat 1620gatacatttg atgaaactta tgctgccacg attggaattg attttttatc gaaaacaatg 1680tatttagaag aaggtaaaac cattagatta caattatggg atactgccgg acaagaaaga 1740tttcgatcat taataccttc atatattaga gattctcatg ttgcagtaat atgttatgat 1800ataaccaata aaaaatcatt tgataatctt gataaatgga ttaaagatgt taaattagaa 1860cgaggtgatg atgtaataat agtattagtc ggtaataaac tggatttagc tagtgataaa 1920cgacaagtta gtttagatga tgttgaaaat ttacaaatta aaattggtgc taaatttttc 1980attgaaactt caactaaagc aaatcataat gttaaattat tatttaaaaa aattgctcaa 2040tcattacctg attttaatca agattccaat gataaatcaa atgataataa taataataat 2100aataataatc aactggaaac tattgatata actattgata atactgcacc aaatcctcaa 2160ggtaccagca catgttgtta gactagaatc ttagtgtaag aactaataaa aaaacagagc 2220aatgggtaga taatattcta agtatattaa cagttcatac aacaacaacc cacacacaca 2280catatatata ctatatatat atatcactta tttaatcaat tagatcaaat tcccaatttg 2340ccgactgtat acgaagtttt aatttatcta atctctcatt cattttcttt cttgtcttgt 2400catatgcaac cacgtcaaca tgtgacacat cagcaatctt cttggtcgaa tacagttttg 2460taataatacc attatgaatt atatcatctg tcacattgat ttgtctttct aaatcatcaa 2520tatcagcaat agcttgtgtc attgatcgaa gtggttcttc attagactta ttatcattat 2580cataggcaaa ctttatttca ttattggttc gttgtatttt aatagttata tcagtaatta 2640atttatcaat ttcttcatat tgttcaaata attcattagg atcaaatgga gggttttcgc 2700cagtttgagc ttgacaacat ttaagaatta atgatttaag ttcaccagca tctctttcca 2760agttcttttt taaatttaat gcttcagcta atttcatctt ctctcacaaa atataatata 2820actcaattat tggttgtaag attatataga ataaagtata tgaaaatgaa aaaaaaatgg 2880ggtgagaggt aaatgtatcc gaatttataa ttctgttgat agcggagaaa agtataattt 2940tatttttttt ttggtagttc ggttgtagtt cctctttgct ttattcccct atgcaccctg 3000gcatacacaa aagtcaattg attgctttct cttgttaagg cttttggggt tggggttgga 3060gtaattgtcg ttgttgttgt tgctgttcct gatacagtgg aatagagata tgactaattg 3120gtattggtat gtttatgttt acacaacaca tatgagtcaa cgaaaaatca attggcttga 3180tctgacttct cctggaacta aattctataa tttcatcaac aattgtaggc aaatgtagac 3240aaatgttgtg gtttcgtcta gctcaatata accatcaagg tttgttaagc ctccttcctt 3300tattattttt gcctcttgaa aggcattttt gatgtaacaa agtgattcta caattgttgc 3360gagcaaatta ttggcaaaca tcttttgtga aagaatcata accttccatt cgtttgttcg 3420ttttgttagc tcattggctg atgggttctt tagttgctat gaatactgct gctctgtttt 3480caaaatcctt ttgttgggaa ggttctaccg attgaggttt aacttgtatt atcgtgtaag 3540tgtgttcctg actccgaatt tttgtctata aatagaccta gaaaagttca ctttttttca 3600aatttttttt tattcccttt ttcttttttc taatcctcat taacaaatca tattcaaaca 3660aatcaatcat tttatgcatt gagtcgtatt aattgttgtt tgttggttat agcttgttgg 3720ttgattgatt ggttggttgg tagtataaac attttcatta ctctaatggc ctcctcagta 3780aagttggcta cggcacttaa acaacgtgct atattgacaa aagaattgtc tgaattagat 3840gataaaatac aatcttcatt gattctgcaa gttggtatga aaaaaatcaa tgatccagat 3900aaattgtatt tagattatgt tgctaaatct caagaattgg ctaaattggt atcatcaata 3960aattatacta ataatataac tccaattgaa cttgatttga caatgggaaa gtatgataat 4020actataaaaa caattaatga tgcattaatt tgtcgagacc gaatatttaa aaaattacaa 4080tttgtgaaaa aaatatcaac agcaggtaaa gaacaaccat tagattccaa agatgaaatt 4140aaatttgtat catttattga tgttgataaa tatgatactt tggcccaaga attaaatact 4200caatttgaga atttgaattt gaaattacaa gaaataaatt ggcaagttga tcttgttgag 4260atataaaaag gatagtggtg ctggatcgcc attgataata ttctttactt gttactttat 4320gtaaaaggat ttaaaaaata ttgttggtac tactcgtttc ctccctccca aatcgaataa 4380tagaactata gaaccatatc ccccctataa ttattttatc tgattttatt agttataaag 4440tacaaatcta ttatcaattg ttttattatt tagtattttc ctccaaagtt ttgaactttt 4500gttttttatg gttctagttc tttattcttg tttttgggga tttagggttg ccgcttgatt 4560tgttgaactt taattgatgc tttgtttagg catagtaatc aagaaaagga agataatgaa 4620agggtaggga atgagtagga gggcgggttc ggggacaata tacatgtata gttacgtaca 4680ttaatgtaaa tatattctta aaattcctag tttgtaaatt aattgatggt gttgttgtct 4740ttgtattttt aaagtattca aaaattttga gtcaatttcg ttaccaaatc ttaatgaata 4800gtaacacgtc taaccaaatt tcaacaaaaa gtttcatacg accaacaact tatatgcttt 4860tcagtatgta tatatcttcc atatttttat ttgtatatga ttgaattgat aattgtaata 4920gagttaaaag aatgaagaag aagaagaagt gggtttttgc aaccaacaga acagttaggt 4980tattcttgtg tacacgacca gatcaaatat gtatgtgaga gagagacgga aatagaattt 5040tctggaaaga aaaaaaaaaa aaaatttcct tcctgttttt ctctcgcccc gtgtgggtgg 5100gtctctctca ctgttgtgta attcgtacca acaattccgg agccaaattt ctttcacc 5158 12968 DNA Candida albicans 12 cttctctcac aaaatataat ataactcaat tattggttgtaagattatat agaataaagt 60 atatgaaaat gaaaaaaaaa tggggtgaga ggtaaatgtatccgaattta taattctgtt 120 gatagcggag aaaagtataa ttttattttt tttttggtagttcggttgta gttcctcttt 180 gctttattcc cctatgcacc ctggcataca caaaagtcaattgattgctt tctcttgtta 240 aggcttttgg ggttggggtt ggagtaattg tcgttgttgttgttgctgtt cctgatacag 300 tggaatagag atatgactaa ttggtattgg tatgtttatgtttacacaac acatatgagt 360 caacgaaaaa tcaattggct tgatctgact tctcctggaactaaattcta taatttcatc 420 aacaattgta ggcaaatgta gacaaatgtt gtggtttcgtctagctcaat ataaccatca 480 aggtttgtta agcctccttc ctttattatt tttgcctcttgaaaggcatt tttgatgtaa 540 caaagtgatt ctacaattgt tgcgagcaaa ttattggcaaacatcttttg tgaaagaatc 600 ataaccttcc attcgtttgt tcgttttgtt agctcattggctgatgggtt ctttagttgc 660 tatgaatact gctgctctgt tttcaaaatc cttttgttgggaaggttcta ccgattgagg 720 tttaacttgt attatcgtgt aagtgtgttc ctgactccgaatttttgtct ataaatagac 780 ctagaaaagt tcactttttt tcaaattttt ttttattccctttttctttt ttctaatcct 840 cattaacaaa tcatattcaa acaaatcaat cattttatgcattgagtcgt attaattgtt 900 gtttgttggt tatagcttgt tggttgattg attggttggttggtagtata aacattttca 960 ttactcta 968 13 456 DNA Candida albicans 13atgtccgacg aaagaacttt tattgctatc aaaccagacg gtgttcaaag aggtttaatc 60tcatctatct tgggtagatt tgaacaaaga ggtttcaaat tagttggtat taaattggtt 120caaccaactg aatctttatt gagaactcat tatgaagatt tacaatctaa accatttttc 180ccatctttat tatcttatat gttatccggt ccagtcttag ctactgtttg ggaaggtaaa 240gatgttgtta aacaaggtag agccattttg ggtgctacta acccattaca atctgctcca 300ggtaccatca gaggtgattt tgccattgat atgggtagaa acgtttgtca tggttctgat 360tctgttgaat ctgctaacaa agaaattgac ttgtggttca agaaagaaga attggttgaa 420tataaaccag ctttgttcgg ttggatctac gaataa 456 14 151 PRT Candida albicans14 Met Ser Asp Glu Arg Thr Phe Ile Ala Ile Lys Pro Asp Gly Val Gln 1 510 15 Arg Gly Leu Ile Ser Ser Ile Leu Gly Arg Phe Glu Gln Arg Gly Phe 2025 30 Lys Leu Val Gly Ile Lys Leu Val Gln Pro Thr Glu Ser Leu Leu Arg 3540 45 Thr His Tyr Glu Asp Leu Gln Ser Lys Pro Phe Phe Pro Ser Leu Leu 5055 60 Ser Tyr Met Leu Ser Gly Pro Val Leu Ala Thr Val Trp Glu Gly Lys 6570 75 80 Asp Val Val Lys Gln Gly Arg Ala Ile Leu Gly Ala Thr Asn Pro Leu85 90 95 Gln Ser Ala Pro Gly Thr Ile Arg Gly Asp Phe Ala Ile Asp Met Gly100 105 110 Arg Asn Val Cys His Gly Ser Asp Ser Val Glu Ser Ala Asn LysGlu 115 120 125 Ile Asp Leu Trp Phe Lys Lys Glu Glu Leu Val Glu Tyr LysPro Ala 130 135 140 Leu Phe Gly Trp Ile Tyr Glu 145 150 15 486 DNACandida albicans 15 atgtctcaat tttacgaatt agctccaaaa gacgccaaaggtgaaccata cccatttgaa 60 caattgaaag ggaaagttgt ccttatcgtc aatgttgcttccaaatgtgg attcactcct 120 caatacaagg gtttagaaga attgaataag aaatttgctgatcaaccagt acaaatcttg 180 ggtttcccat gtaatcaatt tggccaccaa gaaccaggtagtaacgaaga aattggatca 240 ttctgttcat tgaactacgg tgttacattc ccagtcttggataaaattga agtcaatggt 300 gacaataccg atccagttta taaatatttg aaatcacaaaagagtggtgt tttgggattg 360 accagaatta aatggaattt tgaaaaattc ttgattgaccaaaatggtaa agttattgaa 420 agattcagtt cattgactag tccagaaagt atcggtaccaagattgaaga attgttgaag 480 aaataa 486 16 161 PRT Candida albicans 16 MetSer Gln Phe Tyr Glu Leu Ala Pro Lys Asp Ala Lys Gly Glu Pro 1 5 10 15Tyr Pro Phe Glu Gln Leu Lys Gly Lys Val Val Leu Ile Val Asn Val 20 25 30Ala Ser Lys Cys Gly Phe Thr Pro Gln Tyr Lys Gly Leu Glu Glu Leu 35 40 45Asn Lys Lys Phe Ala Asp Gln Pro Val Gln Ile Leu Gly Phe Pro Cys 50 55 60Asn Gln Phe Gly His Gln Glu Pro Gly Ser Asn Glu Glu Ile Gly Ser 65 70 7580 Phe Cys Ser Leu Asn Tyr Gly Val Thr Phe Pro Val Leu Asp Lys Ile 85 9095 Glu Val Asn Gly Asp Asn Thr Asp Pro Val Tyr Lys Tyr Leu Lys Ser 100105 110 Gln Lys Ser Gly Val Leu Gly Leu Thr Arg Ile Lys Trp Asn Phe Glu115 120 125 Lys Phe Leu Ile Asp Gln Asn Gly Lys Val Ile Glu Arg Phe SerSer 130 135 140 Leu Thr Ser Pro Glu Ser Ile Gly Thr Lys Ile Glu Glu LeuLeu Lys 145 150 155 160 Lys 17 1080 DNA Candida albicans 17 atggctcctccagcagtttt aagtaaatcc ggtgttatct acggtaaaga cgtcaaagac 60 ttgtttgactatgctcaaga aaaaggtttt gccattccag ctatcaatgt cacttcatcc 120 tcaactgttgttgctgcttt agaagctgcc agagacaaca aggctccaat catcttgcaa 180 acttctcaaggtggtgctgc ctactttgcc ggtaaaggtg tcgacaacaa agatcaagct 240 gcttccattgctggttcaat tgctgccgct cactacatta gagccattgc tccaacttat 300 ggtatcccagttgttttaca cactgatcac tgtgccaaaa aattattgcc atggtttgat 360 ggtatgttgaaagccgatga agaattcttt gctaagaccg gtactccatt gttctcatcc 420 cacatgttggatttatctga agaaaccgat gacgaaaaca ttgctacttg tgccaaatat 480 ttcgaaagaatggctaaaat gggtcaatgg ttagaaatgg aaattggtat cactggtggt 540 gaagaagatggtgtcaacaa cgaacacgtt gaaaaagatg ctttatacac ttctccagaa 600 actgttttcgctgtctacga atctttacac aagatttctc caaacttttc tattgctgct 660 gcttttggtaacgtccacgg tgtttacaaa ccaggtaatg tgcaattgag accagaaatc 720 ttgggtgaccaccaagttta cgctaagaaa caaattggta ctgatgctaa acacccatta 780 tacttggttttccacggtgg ttctggttct actcaagaag aattcaacac tgctatcaag 840 aatggtgttgtcaaggtcaa cttggacact gattgtcaat acgcttactt gactggtatc 900 agagattacgtcaccaacaa gattgaatac ttgaaagcac cagttggtaa cccagaaggt 960 gctgacaaaccaaacaagaa atactttgac ccaagagtct gggttagaga aggtgaaaag 1020 accatgtccaagagaattgc tgaagctttg gatattttcc acaccaaagg acaattgtaa 1080 18 359 PRTCandida albicans 18 Met Ala Pro Pro Ala Val Leu Ser Lys Ser Gly Val IleTyr Gly Lys 1 5 10 15 Asp Val Lys Asp Leu Phe Asp Tyr Ala Gln Glu LysGly Phe Ala Ile 20 25 30 Pro Ala Ile Asn Val Thr Ser Ser Ser Thr Val ValAla Ala Leu Glu 35 40 45 Ala Ala Arg Asp Asn Lys Ala Pro Ile Ile Leu GlnThr Ser Gln Gly 50 55 60 Gly Ala Ala Tyr Phe Ala Gly Lys Gly Val Asp AsnLys Asp Gln Ala 65 70 75 80 Ala Ser Ile Ala Gly Ser Ile Ala Ala Ala HisTyr Ile Arg Ala Ile 85 90 95 Ala Pro Thr Tyr Gly Ile Pro Val Val Leu HisThr Asp His Cys Ala 100 105 110 Lys Lys Leu Leu Pro Trp Phe Asp Gly MetLeu Lys Ala Asp Glu Glu 115 120 125 Phe Phe Ala Lys Thr Gly Thr Pro LeuPhe Ser Ser His Met Leu Asp 130 135 140 Leu Ser Glu Glu Thr Asp Asp GluAsn Ile Ala Thr Cys Ala Lys Tyr 145 150 155 160 Phe Glu Arg Met Ala LysMet Gly Gln Trp Leu Glu Met Glu Ile Gly 165 170 175 Ile Thr Gly Gly GluGlu Asp Gly Val Asn Asn Glu His Val Glu Lys 180 185 190 Asp Ala Leu TyrThr Ser Pro Glu Thr Val Phe Ala Val Tyr Glu Ser 195 200 205 Leu His LysIle Ser Pro Asn Phe Ser Ile Ala Ala Ala Phe Gly Asn 210 215 220 Val HisGly Val Tyr Lys Pro Gly Asn Val Gln Leu Arg Pro Glu Ile 225 230 235 240Leu Gly Asp His Gln Val Tyr Ala Lys Lys Gln Ile Gly Thr Asp Ala 245 250255 Lys His Pro Leu Tyr Leu Val Phe His Gly Gly Ser Gly Ser Thr Gln 260265 270 Glu Glu Phe Asn Thr Ala Ile Lys Asn Gly Val Val Lys Val Asn Leu275 280 285 Asp Thr Asp Cys Gln Tyr Ala Tyr Leu Thr Gly Ile Arg Asp TyrVal 290 295 300 Thr Asn Lys Ile Glu Tyr Leu Lys Ala Pro Val Gly Asn ProGlu Gly 305 310 315 320 Ala Asp Lys Pro Asn Lys Lys Tyr Phe Asp Pro ArgVal Trp Val Arg 325 330 335 Glu Gly Glu Lys Thr Met Ser Lys Arg Ile AlaGlu Ala Leu Asp Ile 340 345 350 Phe His Thr Lys Gly Gln Leu 355

1. Nucleotide chip including a solid substrate and at least onenucleotide sequence fixed thereon, which is suitable for theidentification and transcription of a gene coding a hyphen-specificprotein selected from the group consisting of: (a) a nucleotide sequencedefined in SEQ ID No. 1, 2, 3, 4, 12, 13, 15 or 17, or a complementarystrand or part thereof, (b) a nucleotide sequence coding the amino acidsequence defined in SEQ ID No. 5, 6, 7, 8, 14, 16 or 18, or acomplementary strand or part thereof, (c) a nucleotide sequence whichhybridizes with one of the nucleotide sequences named in (a) or (b). 2.Nucleotide chip according to claim 1 wherein the nucleotide sequence isa DNA, RNA, or PNA sequence.
 3. Protein chip including a solid substrateand at least one hyphen-specific protein fixed thereon selected from thegroup consisting of: (a) a protein with an amino acid sequence definedin SEQ ID No. 5, 6, 7, 8, 14, 16 or 18, or a fragment thereof, and (b) aprotein with an amino acid sequence which has a sequence identity of atleast 80% to one of the amino acid sequences defined in SEQ ID No. 5, 6,7, 8, 14, 16 or 18, or to a fragment thereof.
 4. Protein chip accordingto claim 3 wherein the fragment of a protein is a protein regionincluding an antigen determinant.
 5. Antibody chip including a solidsubstrate and at least one antibody fixed thereon, which is directedspecifically against a protein with the amino acid sequence representedin SEQ ID 5, 6, 7, 8, 14, 16 or
 18. 6. Antibody chip according to claim5 wherein the antibody is a monoclonal, polyclonal, and/or modifiedantibody.
 7. Antibody chip including a solid substrate and at least oneantibody fixed thereon, which is directed specifically against anantibody against a protein with the amino acid sequence represented inSEQ ID 5, 6, 7, 8, 14, 16 or
 18. 8. Diagnostic composition containing anucleotide chip according to claim 1 or 2, a protein chip according toclaim 3 or 4, or a nucleotide chip according to one of the claims 5 to7.
 9. Process for diagnosing a disease caused by types of the genusCandida wherein a sample to be tested is brought into contact, in asuitable medium, with a nucleotide chip according to claim 1 or 2, aprotein chip according to claim 3 or 4, or an antibody chip according toone of the claims 5 to 7 and an interaction between the sample to betested and at least one of said chips is detected.
 10. Process forfinding and identifying substances which are therapeutically effectiveagainst Candida-caused diseases wherein a substance to be tested isbrought, in a suitable medium, into contact with a nucleotide chipaccording to claim 1 or 2, a protein chip according to claim 3 or 4, oran antibody chip according to one of the claims 5 to 7 and aninteraction between the sample to be tested and at least one of saidchips is detected.